Wnt publications

Roles of nAChR and Wnt signaling in intestinal stem cell function and inflammation.

Journal: International immunopharmacology


Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that signal using endogenous acetylcholine (ACh) and the agonist, nicotine. The nAChR signaling pathway is a central regulator of physiological homeostasis in the central and peripheral nervous systems. The receptors are expressed not only in the nervous system, but also play a pivotal role in regulation of epithelial cell growth, migration, differentiation, and inflammation processes in various mammalian non-neuronal cells. In the intestine, the Wnt signaling pathway plays a central role in the epithelium and is a principal regulator of intestinal stem cell (ISC) identity and proliferation. Since Wnt signaling was first described more than 40 years ago in ISCs, large amounts of scientific evidence have demonstrated remarkable long-term self-renewal capacity of ISCs. Intestinal organoids are commonly used for studying ISC biology and intestinal pathophysiology. The contribution of non-neuronal nAChR signaling to Wnt signaling in the intestine has received less attention. Experiments using cultured intestinal organoids that lack nerve and immune cells were performed. Endogenous ACh is synthesized in the intestinal epithelium and drives organoid growth and differentiation through activation of nAChR signaling. Furthermore, nAChR signaling is coordinated with Wnt signaling for regulation of ISC function. Elucidating the mechanism of the coordinated activities of nAChR and Wnt signaling in the intestine provides new insight into epithelial homeostasis, and may be of particular relevance in inflammatory bowel diseases such as ulcerative colitis and Crohn’s disease.
Toshio Takahashi

Signaling pathways that regulate Trypanosoma cruzi infection and immune response.

Journal: Biochimica et biophysica acta. Molecular basis of diseaseJanuar/31/2020
Current understanding of key cellular pathways, which are activated by the interaction between T. cruzi and host immunity, is crucial for controlling T. cruzi infection and also for limiting the development of the immunopathological symptoms of Chagas´ disease. Here, we focus on recent advances in the knowledge of modulation of innate receptors such as TLRs and NLRs, especially NLRP3, by T. cruzi in different cells of the immune system. On the other hand, the modulation of macrophage activation may be instrumental in allowing parasite persistence and long-term host survival. In this sense, we discuss the importance of the metabolism of two amino acids: L-arginine and tryptophan, and evaluate the role of iNOS, arginase and IDO enzymes in the regulation of innate and adaptive immune response during this infection; and, finally, we also discuss how T. cruzi exploits the AhR, mTOR and Wnt signaling pathways to promote their intracellular replication in macrophages, thus evading the host’s immune response.
Fabio Cerbán; Cinthia Stempin; Ximena Volpini; Antonio Silva; Susana Gea; Claudia Motran

Wnt/beta-catenin and PI3K/Akt/mtor Signaling Pathways in Glioblastoma: Two main targets for drug design: A Review.

Journal: Current pharmaceutical designJanuar/31/2020
Glioblastoma (GBM) is the most common and malignant astrocytic glioma, accounting for about 90% of all brain tumors with poor prognosis. Despite recent advances in understanding molecular mechanisms of oncogenesis and the improved neuroimaging technologies, surgery and adjuvant treatments, the clinical prognosis of patients with GBM remains persistently unfavorable. The signaling pathways and the regulation of growth factors of glioblastoma cells are very abnormal. The various signaling pathways have been suggested to be involved in cellular proliferation, invasion and glioma metastasis. The Wnt signaling pathway with its pleiotropic functions in neurogenesis and stem cell proliferation are implicated in various human cancers, including glioma. In addition, the PI3K/Akt/mTOR pathway is closely related to growth, metabolism, survival, angiogenesis, autophagy, and chemotherapy resistance of GBM. Understanding the mechanisms of GBM’s invasion, represented by invasion and migration, is an important tool in designing effective therapeutic interventions. This review will investigate two main signaling pathways in GBM: PI3K/Akt/mTOR and Wnt/beta-catenin signaling pathways.
Seyed Shahcheraghi; Venant Tchokonte-Nana; Marzieh Lotfi; Malihe Lotfi; Ahmad Ghorbani; HamidReza Sadeghnia

Wnt signaling associated small molecules improve the viability of pPSCs in a PI3K/Akt pathway dependent way.

Journal: Journal of cellular physiologyJanuar/31/2020
Although we have obtained porcine pluripotent stem cell lines (pPSCs) from blastocysts, the cells exhibit flat clonal morphology and do not support single-cell passage. There is massive cell death after cell dissociation, and the efficiency of single-cell colony is generally ≤10%. In a recent study, we got a new pPSCs using two Wnt signaling pathway regulators CHIR99021 and XAV939. This cell had strong biological viability, small-domed morphology, and its cloning efficiency after dissociation was 80-90%. The CH/XAV-treated cells expressed elevated levels of pluripotent genes, and possessed differentiation abilities both in vitro and in vivo, proven by the formation of embryonic bodies and teratomas with three germ layers. Furthermore, we found that the combinative use of CHIR99021 and XAV939 resulted in β-catenin-maintained expression in the cytoplasm but not translocation to the nuclei for WNT/TCF activation. In the meanwhile, E-cadherin located on the cell membrane, thereby activated the PI3K/Akt signaling pathway to enhance the pluripotency of the cells. Our study obtained new pPSCs, which were even closer to the naïve state with only two small molecule inhibitors, and the improved pluripotency of pPSCs could facilitate transgenic manipulation and regenerative medicine research. Besides, our study casted a light on the understanding of pPSCs and the derivation of authentic porcine embryonic stem cells.
Yan Li; Shuang Wu; Xuechun Li; Shimeng Guo; Zhuang Cai; Zhi Yin+2 authors

Integrated analysis of mRNA-miRNA expression in Tilapia infected with Tilapia lake virus (TiLV) and identifies primarily immuneresponse genes.

Journal: Fish & shellfish immunologyJanuar/31/2020
We investigated differential gene expression in Tilapia infected with the Tilapia Lake virus (TiLV).We used high-throughput sequencing to identify mRNAs and miRNAs involved in TiLV infection progression We identified 25,359 differentially expressed genes that included 863 new genes. We identified 1770, 4142 and 4947 differently expressed genes comparing non-infected controls with 24 and 120 h infections and between the infected groups, respectively. These genes were enriched to 291 GO terms and 62 KEGG pathways and included immune system progress and virion genes. High-throughput miRNA sequencing identified 316 conserved miRNAs, 525 known miRNAs and 592 novel miRNAs. Furthermore, 138, 198 and 153 differently expressed miRNAs were found between the 3 groups listed above, respectively. Target prediction revealed numerous genes including erythropoietin isoform X2, double-stranded RNA-specific adenosine deaminase isoform X1, bone morphogenetic protein 4 and tapasin-related protein that are involved in immune responsiveness. Moreover, these target genes overlapped with differentially expressed mRNAs obtained from RNA-seq. These target genes were significantly enriched to GO terms and KEGG pathways including immune system progress, virion and Wnt signaling pathways. Expression patterns of differentially expressed mRNA and miRNAs were validated in 20 mRNA and 19 miRNAs by qRT-PCR. We also were able to construct a miRNA-mRNA target network that can further understand the molecular mechanisms on the pathogenesis of TiLV and guide future research in developing effective agents and strategies to combat TiLV infections in Tilapia.
Yingying Wang; Qing Wang; Yingying Li; Jiyuan Yin; Yan Ren; Cunbin Shi+3 authors

Angiopoietin-like protein 8 (betatrophin) may inhibit hepatocellular carcinoma through suppressing of the Wnt signaling pathway.

Journal: Iranian journal of basic medical sciencesJanuar/30/2020
Hepatocellular carcinoma (HCC) is one of the leading fatal neoplasms and the most common primary liver malignancy worldwide. Peptide hormone ANGPTL8 (betatrophin) may act as an important regulator in HCC development through the Wnt/β-catenin pathway. We aimed to evaluate the effects of recombinant ANGPTL8 on Wnt/β-catenin signaling in human liver carcinoma cells (HepG2) and their viability.The expression of ANGPTL8 was conducted in the pET-21b-E. coli Bl21 (DE3) system and the produced peptide was purified. HepG2 cells were treated with different concentrations of ANGPTL8 (25, 50, 100, 150, 200, and 250 ng/ml) for 24, 48, and 72 hr. MTT assay was performed to detect the viability of treated cells, and apoptotic induction by ANGPTL8 was measured by flow cytometry assay. Finally, using qRT-PCR the mRNA levels of Wnt signaling modulators WIF-1 and β-catenin were evaluated in treated cells.
MTT assay showed that ANGPTL8 inhibits proliferation of HepG2 cells moderately in a time-independent manner. The highest concentration of the ANGPTL8, 250 ng/ml, reduced cell proliferation after 24, 48, and 72 hr similarly about 30%. In the same concentration of ANGPTL8, after 24 hr of treatment flow cytometry assay revealed a mild increase in early and late apoptosis up to 7.7 and 14.3%, respectively. The qRT-PCR showed that in a concentration-dependent and time-independent fashion, the expression of WIF-1 and β-catenin genes respectively increased and decreased significantly (P<0.05).
Our findings suggest that ANGPTL8 may act as a moderate suppressor against HCC cell proliferation possibly via affecting Wnt signaling modulators.
Nastaran Monzavi; Seyed Zargar; Nematollah Gheibi; Mahdi Azad; Babak Rahmani

The physiological role of Wnt pathway in normal development and cancer.

Journal: Experimental biology and medicine (Maywood, N.J.)Januar/30/2020
Xiang Li; Maria Ortiz; Leszek Kotula

Clinical Implications of Primary Cilia in Skin Cancer.

Journal: Dermatology and therapyJanuar/30/2020
The primary cilium is a cell surface organelle that is an important component of cellular biology. While it was once believed to be a vestigial structure without biologic function, it is now known to have essential roles in critical cellular signaling pathways such as Hedgehog (HH) and Wnt. The HH and Wnt pathways are involved in pathogenesis of basal cell carcinoma and melanoma, respectively, and this knowledge is now beginning to inform therapeutic and diagnostic options for patients. The purpose of this review is to familiarize clinicians with primary cilia biology and how this complex cellular organelle has started to translate into clinical care.
Abrar Choudhury; Neil Neumann; David Raleigh; Ursula Lang
Aberrant non-canonical WNT pathway as key-driver of high-grade serous ovarian cancer development.

Journal: Virchows Archiv : an international journal of pathologyJanuar/29/2020
Gian Zannoni; Giuseppe Angelico; Angela Santoro

VGLL3 is a prognostic biomarker and correlated with clinical pathologic features and immune infiltrates in stomach adenocarcinoma.

Journal: Scientific reportsJanuar/29/2020
Due to its poor clinical outcome, there is an urgent need to identify novel prognostic markers for stomach adenocarcinoma (STAD). Here, we aimed to explore the relationship between VGLL3 expression and clinico-pathological features, dendritic cells, macrophages, and prognosis of STAD. VGLL3 expression levels were significantly associated with histological grade, T stage, and TNM stage. VGLL3 levels and patient’s age were also independent prognostic factors of the clinical outcome of STAD. In addition, VGLL3 was associated with the abundance of macrophages and dendritic cells in tumor infiltrates, of which only VGLL3 and macrophage counts were the independent prognostic factors of immune cell infiltration in the TIMER Database. Extracellular matrix receptor interaction, focal adhesion, pathways in cancer, MAPK, JAK STAT, and WNT signaling pathways were enriched in VGLL3 high-expressing datasets as determined by Gene Set Enrichment Analysis (GSEA), while DNA replication, glyoxylate, and dicarboxylate metabolism, glutathione metabolism, homologous recombination, and glycosylphosphatidylinositol gpi banchor biosynthesis were enriched in VGLL3 low-expressing datasets. Thus, VGLL3 is a novel prognostic biomarker of both the clinical outcome and immune infiltration in STAD, and may therefore be a promising therapeutic target.
Lihua Zhang; Longhai Li; Yong Mao; Dong Hua

Nuclear-localized costimulatory molecule 4-1BBL promotes colon cancer cell proliferation and migration by regulating nuclear Gsk3β, and is linked to the poor outcomes associated with colon cancer.

Journal: Cell cycle (Georgetown, Tex.)Januar/29/2020
Anti-tumor immune response and the prognosis of tumor are the results of competition between stimulatory and inhibitory checkpoints. Except for upregulating inhibitory checkpoints, lowering some immune accelerating molecules to convert an immunostimulatory microenvironment into an immunodormant one through “decelerating the accelerator” might be another effective immune escape pattern. 4-1BBL is a classical transmembrane costimulatory molecule involving in antitumor immune responses. In contrast, we demonstrated that 4-1BBL is predominantly localized in the nuclei of cancer cells in colon cancer specimens and is positively correlated with tumor size, lymph node metastasis, and a lower survival ratio. Furthermore, the nuclear localization of 4-1BBL was also ascertained in vitro. 4-1BBL knockout (KO) arrests the proliferation and impaired the migration and invasion ability of colon cancer cells in vitro and retarded tumor growth in vivo. 4-1BBL KO increased the accumulation of Gsk3β in the nuclei of colon cancer cells and consequently decreased the expression of Wnt pathway target genes and thus alter tumor biological behavior. We hypothesized that unlike membrane-expressed 4-1BBL, which stimulates the 4-1BB signaling of antitumor cytotoxic T cells, the nuclear-localized 4-1BBL could facilitate the malignant behavior of colon cancer cells by circumventing antitumor signaling and driving some key oncotropic signal pathway in the nucleus. Nuclear-localized 4-1BBL might be an indicator of colon cancer malignancy and serve as a promising target of immunotherapy.
Yan Ge; Wei Chen; Xueguang Zhang; Haiyan Wang; Juanjuan Cui; Yue Liu+3 authors

Elevated levels of the secreted wingless agonist R-spondin 3 in preeclamptic pregnancies.

Journal: Journal of hypertensionJanuar/28/2020
Preeclampsia is a syndrome characterized by hypertension and poor placental development. The developmental wingless (Wnt) pathway plays an important role in placental development and we hypothesized that Wnt signaling would be dysregulated in preeclampsia.To elucidate aberrations in the Wnt signaling pathway we conducted a pathway analysis on placental mRNA in late-onset preeclampsia and normal pregnancy from the STORK study [n = 10 in each group, RNA sequencing (RNAseq)] to identify differentially expressed genes. In addition, we compared circulating levels of secreted Wnt agonists and antagonists at term pregnancy and 6 months postpartum from an acute preeclampsia study (preeclampsia n = 34, normal pregnancy n = 61).We found circulating and placental mRNA levels of the secreted Wnt agonist R-spondin 3 (RSPO3) at term elevated in preeclampsia. Increased plasma RSPO3 was associated with high mean arterial pressure. Further, pathway analysis of placental tissue revealed elevated mRNA levels of upstream ligands WNT6 and WNT10A and frizzled receptors 2 and 4 in preeclampsia and downstream activation of the noncanonical Ca/NFAT pathway. Finally, plasma dickkopf 3 was decreased in preeclampsia 6 months postpartum.We identify a potential role for RSPO3 and activation of noncanonical Wnt signaling in preeclampsia.
Thor Ueland; Mette-Elise Estensen; Guro Grindheim; Jens Bollerslev; Tore Henriksen; Pål Aukrust+3 authors

Exploring genetic targets of psoriasis using genome wide association studies (GWAS) for drug repurposing.

Journal: 3 BiotechJanuar/28/2020
Psoriasis is a chronic inflammatory disease causing itching in the body and pain in the joints. Currently, no permanent cure is available at a commercial level for this disease. Genome wide association studies (GWAS) provide a deeper insight that helps in better understanding this disease and further possible cure of this disease. The major goal of the present study is to identify potent genetic targets of psoriasis disease using GWAS approach and identify drugs for repurposing. The methods used include GWAS catalogue, GeneAnalytics, canSAR protein annotation tool, VarElect, Drug bank, Proteomics database, ProTox software. By exploring GWAS catalogue, 126 psoriasis associated genes (PAG) were identified. 68 genes found to be druggable were obtained from canSAR protein annotation tool. Localization results depict that maximum genes are present in cytoplasmic cellular components. The superpathways obtained from GeneAnalytics resulted in involvement of these genes in the immune system, Jak/Stat pathway, Th17 and Wnt pathways. Two genes Interleukin 13 (IL13) and POLI are Food and Drug Administration (FDA) approved targets. Small compounds for these targets were analysed for drug-likeliness, toxicity and mutagenecity properties. The FDA approved drug pandel was found to possess desirable properties. The medications used for psoriasis causes mild to severe side effects and does not work well always. Hence we propose drug repurposing strategy to use existing drugs for new therapies. Therefore, the drug pandel could be explored further and repurposed to treat psoriasis.
Harshit Nanda; Nirmaladevi Ponnusamy; Rajasree Odumpatta; Jeyaraman Jeyakanthan; Arumugam Mohanapriya

Cytokeratin 5 alters β-catenin dynamics in breast cancer cells.

Journal: OncogeneJanuar/28/2020
Estrogen receptor (ER) positive breast cancers often contain subpopulations of cells that express the intermediate filament protein cytokeratin 5 (CK5). CK5+ cells are enriched in cancer stem cell (CSC) properties, can be induced by progestins, and predict poor prognosis in ER+ breast cancer. We established through CK5 knockout and overexpression in ER+ breast cancer cell lines that CK5 is important for tumorsphere formation, prompting us to speculate that CK5 has regulatory activity in CSCs. To interrogate CK5 interacting proteins that may be functionally cooperative, we performed immunoprecipitation-mass spectrometry for CK5 in ER+ breast cancer cells. Focusing on proteins with signaling activity, we identified β-catenin, a key transcription factor of the Wnt signaling pathway and cell adhesion molecule, as a CK5 interactor, which we confirmed by co-immunoprecipitation in several breast cancer models. We interrogated the dual functions of β-catenin in relation to CK5. Knockout or knockdown of CK5 ablated β-catenin transcriptional activity in response to progestins and Wnt stimuli. Conversely, CK5 induced by progestins or overexpression was sufficient to promote the loss of β-catenin at the cell membrane and total E-cadherin loss. A breast cancer patient-derived xenograft showed similar loss of membrane β-catenin and E-cadherin in CK5+ but not intratumoral CK5- cells and single-cell RNA sequencing found the top enriched pathways in the CK5+ cell cluster were cell junction remodeling and signaling. This report highlights that CK5 actively remodels cell morphology and that blockade of CK5-β-catenin interaction may reverse the detrimental properties of CK5+ breast cancer cells.
Olivia McGinn; Ashley Ward; Lynsey Fettig; Duncan Riley; Joshua Ivie; Kiran Paul+3 authors

A Cell Fate Switch in the Caenorhabditis elegans Seam Cell Lineage Occurs Through Modulation of the Wnt Asymmetry Pathway in Response to Temperature Increase.

Journal: GeneticsJanuar/28/2020
Populations often display consistent developmental phenotypes across individuals despite the inevitable biological stochasticity. Nevertheless, developmental robustness has limits and systems can fail upon change in the environment or the genetic background. We use here the seam cells, a population of epidermal stem cells in Caenorhabditis elegans, to study the influence of temperature change and genetic variation on cell fate. Seam cell development has mostly been studied so far in the lab reference strain (N2), grown at 20° temperature. We demonstrate that an increase in culture temperature to 25°, introduces variability in the wild-type seam cell lineage with a proportion of animals showing an increase in seam cell number. We map this increase to lineage-specific symmetrisation events of normally asymmetric cell divisions at the fourth larval stage, leading to the retention of seam cell fate in both daughter cells. Using genetics and single molecule imaging, we demonstrate that this symmetrisation occurs via changes in the Wnt asymmetry pathway, leading to aberrant Wnt target activation in anterior cell daughters. We find that intrinsic differences in the Wnt asymmetry pathway already exist between seam cells at 20° and this may sensitise cells towards a cell fate switch at increased temperature. Finally, we demonstrate that wild isolates of C. elegans display variation in seam cell sensitivity to increased culture temperature, although their average seam cell number is comparable at 20°. Our results highlight how temperature can modulate cell fate decisions in an invertebrate model of stem cell patterning.
Mark Hintze; Sneha Koneru; Sophie Gilbert; Dimitris Katsanos; Julien Lambert; Michalis Barkoulas

Neuronal Excitability in Epileptogenic Zones Regulated by the Wnt/Β-Catenin Pathway.

Journal: CNS & neurological disorders drug targetsJanuar/28/2020
Epilepsy is a neurological disorder that involves abnormal and recurrent neuronal discharges, producing epileptic seizures. Recently, it has been proposed that the Wnt signaling pathway is essential for central nervous system development and function because it modulates important processes such as hippocampal neurogenesis, synaptic clefting, and mitochondrial regulation. Wnt/β-catenin signaling regulates changes induced by epileptic seizures, including neuronal death. Several genetic studies associate Wnt/β-catenin signaling with neuronal excitability and epileptic activity. Mutations and chromosomal defects underlying syndromic or inherited epileptic seizures have been identified. However, genetic factors underlying the susceptibility of an individual to develop epileptic seizures have not been fully studied yet. In this review, we describe the genes involved in neuronal excitability in epileptogenic zones dependent on the Wnt/β-catenin pathway.
Carmen Rubio; Elisa Taddei; Jorge Acosta; Verónica Custodio; Carlos Paz

Long noncoding RNA expression profiles during the NEL-like 1 protein-induced osteogenic differentiation.

Journal: Journal of cellular physiologyJanuar/27/2020
Long noncoding RNAs (lncRNAs) are important modulators of mesenchymal stem cells (MSCs) in cellular differentiation. However, the regulatory mechanisms of lncRNAs in NEL-like 1 (NELL-1)-induced osteogenic differentiation of human adipose-derived stem cells remain elusive. Expression profiles of lncRNAs and messenger RNAs during NELL-1-induced osteogenesis were obtained using high-throughput sequencing. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes pathway analysis, and gene coexpression networks were performed. We identified 323 statistically differentially expressed lncRNAs during osteogenesis and NELL-1-induced osteogenesis, and three lncRNAs (ENST00000602964, ENST00000326734, and TCONS_00006792) were identified as core regulators. Hedgehog pathway markers, including IHH and GLI1, were downregulated, while the antagonists of this pathway (GLI3 and HHIP) were upregulated during NELL-1-induced osteogenesis. In this process, the antagonist of Wnt, SFRP1, was downregulated. According to the analysis, we speculated that lncRNAs played important roles in NELL-1-induced osteogenesis via the crosstalk between Hedgehog and Wnt pathways.
Kai Xia; Xiao Cen; Liyuan Yu; Xinqi Huang; Wentian Sun; Zhihe Zhao; Jun Liu

MiR-128 promotes osteogenic differentiation of bone marrow mesenchymal stem cells in rat by targeting DKK2.

Journal: Bioscience reportsJanuar/27/2020
Bone loss caused by inflammatory disease, such as peri-implantitis, poses a great challenge to clinicians for restoration. Emerging evidence indicates that microRNAs (miRNAs) are indispensable regulators of bone growth, development, and formation. In this study, we found that microRNA-128 (miR-128) was differentially upregulated during the osteogenic differentiation of rat bone marrow stem cells (rBMSCs). Overexpression of miR-128 promoted osteogenic differentiation of rBMSCs by upregulating alkaline phosphatase (ALP), matrix mineralization, mRNA and protein levels of osteogenic makers (e.g., RUNX2, BMP-2, and COLIA1), whereas inhibition of miR-128 suppressed osteoblastic differentiation in vitro. Mechanistically, miR-128 directly and functionally targeted Dickkopf2 (DKK2), which is a Wnt signaling pathway antagonist, and enhanced Wnt / β-catenin signaling activity. Furthermore, the positive effect of miR-128 on osteogenic differentiation was apparently abrogated by DKK2 overexpression. Collectively, these results indicate that miR-128 promotes osteogenic differentiation of rBMSCs by targeting DKK2, which may provide a promising approach to the treatment of peri-implantitis.
Can Wang; Xianghe Qiao; Zhuang Zhang; Chunjie Li

A novel risk score model for stomach adenocarcinoma based on the expression levels of 10 genes.

Journal: Oncology lettersJanuar/22/2020
Stomach adenocarcinoma (STAD) accounts for 95% of cases of malignant gastric cancer, which is the third leading cause of cancer-associated mortality worldwide. The pathogenesis and effective diagnosis of STAD have become popular topics for research in the previous decade. In the present study, high-throughput RNA sequencing expression profiles and clinical data from patients with STAD were obtained from The Cancer Genome Atlas database and were used as a training dataset to screen differentially expressed genes (DEGs). Prognostic DEGs were identified using univariate Cox regression analysis and were further screened by the least absolute shrinkage and selection operator regularization regression algorithm. The resulting genes were used to construct a risk score model, the validation and effectiveness evaluation of which were performed on an independent dataset downloaded from the Gene Expression Omnibus database. Stratified and functional pathway (gene set enrichment) analyses were performed on groups with different estimated prognosis. A total of 92 genes significantly associated with STAD prognosis were obtained by univariate Cox regression analysis, and 10 prognosis-associated DEGs; hemoglobin b, chromosome 4 open reading frame 48, Dickkopf WNT signaling pathway inhibitor 1, coagulation factor V, serpin family E member 1, transmembrane protein 200A, NADPH oxidase organizer 1, C-X-C motif chemokine ligand 3, mannosidase endo-α-like and tripartite motif-containing 31; were selected for the development of the risk score model. The reliability of this prognostic method was verified using a validation set, and the results of multivariate Cox analysis indicated that the risk score may serve as an independent prognostic factor. In functional DEG analysis, eight Kyoto Encyclopedia of Genes and Genomes pathways were identified to be significantly associated with STAD risk factors. Thus, the 10-gene risk score model established in the present study was regarded as credible. This risk assessment tool may help identify patients with a high risk of STAD, and the proposed prognostic mRNAs may be useful in elucidating STAD pathogenesis.
Encui Guan; Feng Tian; Zhaoxia Liu

Targeted molecular profiling of genetic alterations in colorectal cancer using next-generation sequencing.
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Journal: Oncology lettersJanuar/22/2020
Colorectal cancer (CRC) is a major contributor to cancer-associated mortality in China and remains a vast challenge worldwide. Although the genetic basis of CRC has been investigated, the uncommonly mutated genes in CRC remain unknown, in particular in the Asian population. In the present study, targeted region sequencing on 22 CRC and 10 paired non-cancerous tissues was performed to determine the genetic pattern of CRC samples in the Chinese population. Driver genes were detected by three distinct softwares, including MutSigCV, oncodriveFM and iCAGES. A total of 1,335 reliable somatic mutations were identified in tumour samples compared with normal samples. Furthermore, mismatch repair (MMR) mutant patients presented significantly higher mutation density compared with MMR wild-type patients. The results from MutSigCV, oncodriveFM and iCAGES analyses simultaneously detected 29 unique driver genes. In addition, the genes APC regulator of WNT signaling pathway, SMAD family member 4, neurofibromin 1, AT-rich interaction domain 5B and nuclear receptor corepressor 1 were the top five most frequently mutated genes in CRC samples, with mutation rates of 68, 36, 36, 32 and 27%, respectively. The findings from the present study may therefore serve as a basis for future investigation on the diagnosis and oncogenesis of CRC.
Jia Luo; Shengjun Zhang; Meihua Tan; Jia Li; Huadong Xu; Yanfei Tan; Yue Huang

Dysregulation of the Wnt signaling pathway and synovial stem cell dysfunction in osteoarthritis development.

Journal: Stem cells and developmentJanuar/22/2020
Stem cell dysfunction and failure have been found in joints afflicted by osteoarthritis (OA). However, the exact factors in the OA microenvironment that impair stem cell functions and the role of stem cell dysfunction in OA development have not been fully clarified. In this study, we evaluated the functional status of synovial mesenchymal stem cells (SMSCs) from OA patients and explored the influence of OA-SMSCs on cartilage degradation in a rat model. We then screened 138 Wnt signaling-related genes in the synovium of OA patients, focusing on the effects of 5 WNT ligands on SMSC functions. The OA synovium showed mild hyperplasia, and we found a large number of CD90+/CD105+ stem cells in synovial hyperplasia. The OA-SMSCs revealed a cellular senescence phenotype, with decreased proliferation and chondrogenic capacity, accompanied by enhanced migration, proinflammatory and matrix degradation activities. The intra-articular transplantation of these OA-SMSCs significantly aggravated the degradation and destruction of the articular cartilage. Of 138 Wnt signaling genes, the expression of 86 genes was consistently altered in the OA synovium, among which the increased expression of DVL2, WNT10A and DKK3 was the most marked. In general, we found that canonical Wnt/β-catenin pathways were inhibited in the OA synovium, whereas noncanonical PCP and Wnt/Ca2+ pathways were activated. In vitro, WNT10A had an obvious antisenescence effect on SMSCs. WNT5B significantly inhibited the chondrogenic differentiation of SMSCs, and WNT10A and WNT5A increased the expression of inflammatory cytokines in SMSCs. In a rat model, WNT5A significantly aggravated joint degeneration, whereas WNT10A had a mild protective effect on cartilage integrity. In conclusion, stem cells in the OA synovium were functionally abnormal and promoted the development of OA, while dysregulation of the Wnt signaling pathway revealed a comprehensive influence on SMSC functions and cartilage degradation.
Junjie Huang; Chuanshun Chen; Chi Liang; Pan Luo; Guang Xia; Lina Zhang+4 authors

Long noncoding RNA CCAT2 promotes proliferation and metastasis in non-small cell lung cancer through the Wnt pathway.

Journal: International journal of clinical and experimental pathologyJanuar/22/2020
Colon cancer associated transcript 2 (CCAT2), a long non-coding RNA (lncRNA), was shown to be associated with colon, ovarian and prostate cancer. Recent studies showed CCAT2 was highly expressed in various tumors, including non-small cell lung cancer (NSCLC) and probably being an independent prognostic factor of this disease. However, the physiological and biochemical mechanisms for CCAT2 with NSCLC were still unknown. In this study, we performed the analysis focused on the expression, biological functions and mechanism of CCAT2 in NSCLC and found that CCAT2 was significantly up-regulated in NSCLC tissues compared with corresponding non-cancerous tissues. Knockout of CCAT2 in NSCLC cell lines using lentivirus-mediated RNA interference significantly inhibited the proliferation and metastasis of the NSCLC cells. Most importantly, Wnt/β-catenin pathway was found to be inactivated in the NSCLC cell lines after CCAT2 knockout experiment. These results indicated that CCAT2 maybe serve as an oncogenic lncRNA that promoted proliferation and metastasis of NSCLC and activated the Wnt/β-catenin pathway.
Jijia Li; Yu Liu; Pengfei Li; Yingwei Guo; Yannan Liu; Yi Ren

Transcriptomic analysis of stromal cells from patients with endometrial carcinoma.

Journal: International journal of clinical and experimental pathologyJanuar/22/2020
Tumor microenvironment plays a critical role in cancer pathogenesis. In this study, we performed transcriptomic analysis of stromal cells from patients diagnosed with endometrial carcinoma. Endometrial stromal cells from patients and healthy donors were cultured and total RNA was extracted for RNA integrity examination and gene profiling analysis. Gene ontology (GO) and KEGG analysis were also performed. In this study, we found that, in endometrial stromal cells from endometrial cancer patients, a total of 605 genes were changed (fold change ≥2, p-value <0.05). From these, 275 were up-regulated and 330 were down-regulated genes. In addition, GO analysis showed that the differentially expressed genes (DEGs) were involved in various biological processes including cell adhesion, biological adhesion, bone development and extracellular matrix organization. Furthermore, KEGG analysis of the DEGs identified four pathways including Wnt signaling pathway, cadherin signaling pathway, ECM-receptor interaction, and focal adhesion. Our study identified 605 DEGs in stromal cells from endometrial carcinoma which mapped to a variety of biological processes. These results may contribute to understanding the molecular mechanisms o of endometrial carcinoma pathogenesis.
Min Li; Xiao-Yan Xin; Zhi-Shan Jin; Teng Hua; Hong-Bin Wang; Hong-Bo Wang

Silencing CCAT2 inhibited proliferation and invasion of epithelial ovarian carcinoma cells by regulating Wnt signaling pathway.

Journal: International journal of clinical and experimental pathologyJanuar/22/2020
Long non-coding RNA CCAT2 (colon cancer-associated transcript 2) is dysregulated in varieties of human tumors. However, the role of CCAT2 in epithelial ovarian carcinoma (EOC) is not yet known clearly. The aim of this study is to investigate the effects of CCAT2 on proliferation and invasion of EOC cells and the potential mechanisms by which CCAT2 functions. In the present paper, we found that knockdown of CCAT2 impaired cell proliferation and invasion in vitro. Furthermore, we also studied the role of CCAT2 in the modulation of Wnt/β-catenin signaling pathway. Our results showed that knockdown of CCAT2 inhibited the expression of β-catenin and the activity of TCF/LEF (T-cell factor/lymphoid enhancer factor) acting as a key transcription factor of Wnt/β-catenin signaling pathway. In addition, we found that silencing CCAT2 down-regulated the expression of c-MYC and MMP-7. But, that was reversed by the treatment with LiCl (lithium chloride) which could activate canonical Wnt/β-catenin signaling pathway. Taken together, these results indicate that CCAT2 may promote ovarian cancer progression, at least partly, through Wnt/β-catenin signaling pathway. Thus, CCAT2 might represent a novel therapeutic target for ovarian cancer.
Bei-Di Wang; Jing Jiang; Mei-Mei Liu; Ru-Jin Zhuang; Hao Wang; Pei-Ling Li

ROR1 promotes the proliferation of endometrial cancer cells.

Journal: International journal of clinical and experimental pathologyJanuar/22/2020
Endometrial cancer (EC) is the most common gynecological malignant tumor. The canonical Wnt/β-catenin signaling pathway plays a key role in regulating carcinogenesis, and the noncanonical Wnt5a-ROR1 pathway is an important regulator of Wnt signaling. However, the molecular mechanism by which ROR1 influences Wnt signaling in EC is not known. In this study, we found that ROR1 is expressed at higher levels in tumor tissues and blood samples from patients with stage II EC compared with patients with stage I disease. In vitro, human EC cell lines stably overexpressing ROR1 proliferated more rapidly and formed larger colonies than control cells. Consistent with this, overexpression or knockdown of ROR1 increased or decreased, respectively, the percentage of EC cells in M phase of the cell cycle. Elevated levels of ROR1 were associated with increased expression of Wnt5a and of cyclin D1 and c-Myc, two components of the Wnt signaling pathway. Finally, nude mice grew significantly larger tumors after subcutaneous injection of ROR1-overexpressing EC cells compared with control cells. These findings indicate a novel role for ROR1 in promoting EC cell proliferation by upregulating Wnt5a and stimulating the Wnt/β-catenin signaling pathway.
Huilin Zhang; Xiaofang Yan; Jieqi Ke; Yongli Zhang; Chenyun Dai; Mei Zhu+2 authors

MicroRNAs and regulated interaction networks reveal differences between adult and pediatric acute myeloid leukemia.

Journal: International journal of clinical and experimental pathologyJanuar/22/2020
The purpose of this study was to identify featured microRNAs and their regulated network between adult and pediatric acute myeloid leukemia (AML) and find potential utility as biomarkers for diagnosis and treatment of pediatric AML.We downloaded the microRNA expression dataset GSE35320 from Gene Expression Omnibus database and selected expression chips from bone marrow of 71 pediatric AML samples and 6 adulthood AML samples. Differentially expressed microRNAs were identified by Wilcox test. The target genes of these microRNAs were predicted using an integrative method and their functional enrichment analysis was performed using DAVID. Finally, STRING database and Cytoscape software was used to construct and analyze the interaction network.A total of 7 differentially expressed microRNAs were identified and the remarkably up-regulated and down-regulated microRNAs were miR-16 and miR-142-5p which included 323 and 22 predicted target genes, respectively. The target genes of 7 microRNAs were most associated with regulation of cell cycle, p53 signaling pathway, Wnt signaling pathway and neurotrophin signaling pathway. The interaction network of miR-16 target genes was constructed among 354 high confidence interaction pairs. The core genes of the network, such as TP53, BCL2, VEGFA, had a role in prognosis of children with AML.The featured microRNAs and their target genes are significant in the occurrence and development of pediatric AML, which is likely to be important for the identification of therapeutic targets and biomarkers for these patients.
Jing Liu; Lei Zhou; Xiaomin Fu; Chen Xu; Sai Huang; Yan Li+4 authors

Down-regulation of SLC35C1 induces colon cancer through over-activating Wnt pathway.

Journal: Journal of cellular and molecular medicineJanuar/21/2020
The canonical Wnt signalling pathway is a critical pathway involved in the proliferation of cells. It has been well-established that it plays the central role during colorectal carcinogenesis and development. Yet the exact molecular mechanism of how the canonical Wnt pathway is fine-tuned remains elusive. We found that SLC35C1, a GDP-fucose transporter, negatively regulates the Wnt signalling pathway. We show here that SLC35C1 is reduced in all colon cancer by both immunohistochemistry images and TCGA data, whereas β-catenin is increased. Down-regulation of SLC35C1 is also detected by real-time PCR in stage 3 and stage 4 colorectal cancer tissues. Moreover, analysing the TCGA database with cBioPortal reveals the negative correlation of SLC35C1 mRNA level to the expression of β-catenin. Reduced SLC35C1 significantly promotes cell proliferation and colony formation of HEK293 cells. Meanwhile, in HEK293 cells silencing SLC35C1 activates canonical Wnt pathway, whereas overexpressing SLC35C1 inhibits it. Consistently, the reduction of SLC35C1 in HEK293 cells also elevated the mRNA level of Wnt target genes C-myc, Axin2 and Cyclin D1, as well as the secretion of Wnt3a. In conclusion, we identified SLC35C1 as a negative regulator of the Wnt signalling pathway in colon cancer. Decreased SLC35C1 may cause over-activation of Wnt signalling in colorectal cancer.
Minzi Deng; Zhihong Chen; Jieqiong Tan; Heli Liu

Neuroprotective effects of isoquercitrin in diabetic neuropathy via Wnt/β-catenin signaling pathway inhibition.

Journal: BioFactors (Oxford, England)Januar/21/2020
Diabetic neuropathy is a peripheral nervous system disorder affecting both somatic and autonomic components of nervous system. A growing body of evidence have depicted that high glucose levels can induce activation of the Wnt/β-catenin pathway, however there are no studies targeting this pathway in DN. The intent of the present study was to investigate the effects of isoquercitrin (ISQ), a Wnt/β-catenin signaling pathway inhibitor, in diabetic neuropathy. Streptozotocin (50 mg/kg, i.p.) was used to induce diabetes in rats. 6-week diabetic rats were treated intrathecally with ISQ at 10 and 30 μM doses for 3 days. Furthermore, to confirm the results of the intrathecal study, a 2-week intraperitoneal treatment of ISQ was given to diabetic rats. After 6 weeks, diabetic rats developed neuropathy which was evident from reduced thermal and mechanical hyperalgesia thresholds and significant deterioration in motor nerve conduction velocity (MNCV), nerve blood flow (NBF). Sciatic nerves of diabetic neuropathy rats showed increased expression of Wnt pathway proteins namely β-catenin, c-myc and MMP2. Treatment with ISQ, both intrathecally (10 and 30 μM) and intraperitoneally (10 mg/kg), significantly ameliorated the alterations in behavioral pain thresholds and improved functional parameters in diabetic rats. Moreover, ISQ also downregulated the expression of Wnt/β-catenin pathway proteins significantly in diabetic rats as compared to vehicle-treated diabetic rats. Results of the present study suggest the neuroprotective potential of ISQ in the treatment of DN via inhibition of Wnt/β-catenin signaling pathway.
Kahkashan Resham; Pragyanshu Khare; Mahendra Bishnoi; Shyam Sharma

The FGF, TGFβ and WNT axis Modulate Self-renewal of Human SIX2+ Urine Derived Renal Progenitor Cells.

Journal: Scientific reportsJanuar/21/2020
Human urine is a non-invasive source of renal stem cells with regeneration potential. Urine-derived renal progenitor cells were isolated from 10 individuals of both genders and distinct ages. These renal progenitors express pluripotency-associated proteins- TRA-1-60, TRA-1-81, SSEA4, C-KIT and CD133, as well as the renal stem cell markers -SIX2, CITED1, WT1, CD24 and CD106. The transcriptomes of all SIX2+ renal progenitors clustered together, and distinct from the human kidney biopsy-derived epithelial proximal cells (hREPCs). Stimulation of the urine-derived renal progenitor cells (UdRPCs) with the GSK3β-inhibitor (CHIR99021) induced differentiation. Transcriptome and KEGG pathway analysis revealed upregulation of WNT-associated genes- AXIN2, JUN and NKD1. Protein interaction network identified JUN- a downstream target of the WNT pathway in association with STAT3, ATF2 and MAPK1 as a putative negative regulator of self-renewal. Furthermore, like pluripotent stem cells, self-renewal is maintained by FGF2-driven TGFβ-SMAD2/3 pathway. The urine-derived renal progenitor cells and the data presented should lay the foundation for studying nephrogenesis in human.
Md Rahman; Wasco Wruck; Lucas-Sebastian Spitzhorn; Lisa Nguyen; Martina Bohndorf; Soraia Martins+5 authors

Production, purification and characterization of recombinant human R-spondin1 (RSPO1) protein stably expressed in human HEK293 cells.

Journal: BMC biotechnologyJanuar/21/2020
The R-Spondin proteins comprise a family of secreted proteins, known for their important roles in cell proliferation, differentiation and death, by inducing the Wnt pathway. Several studies have demonstrated the importance of RSPOs in regulation of a number of tissue-specific processes, namely: bone formation, skeletal muscle tissue development, proliferation of pancreatic β-cells and intestinal stem cells and even cancer. RSPO1 stands out among RSPOs molecules with respect to its potential therapeutic use, especially in the Regenerative Medicine field, due to its mitogenic activity in stem cells. Here, we generated a recombinant human RSPO1 (rhRSPO1) using the HEK293 cell line, obtaining a purified, characterized and biologically active protein product to be used in Cell Therapy. The hRSPO1 coding sequence was synthesized and subcloned into a mammalian cell expression vector. HEK293 cells were stably co-transfected with the recombinant expression vector containing the hRSPO1 coding sequence and a hygromycin resistance plasmid, selected for hygror and subjected to cell clones isolation.
rhRSPO1 was obtained, in the absence of serum, from culture supernatants of transfected HEK293 cells and purified using a novel purification strategy, involving two sequential chromatographic steps, namely: heparin affinity chromatography, followed by a molecular exclusion chromatography, designed to yield a high purity product. The purified protein was characterized by Western blotting, mass spectrometry and in vitro (C2C12 cells) and in vivo (BALB/c mice) biological activity assays, confirming the structural integrity and biological efficacy of this human cell expression system. Furthermore, rhRSPO1 glycosylation analysis allowed us to describe, for the first time, the glycan composition of this oligosaccharide chain, confirming the presence of an N-glycosylation in residue Asn137 of the polypeptide chain, as previously described. In addition, this analysis revealing the presence of glycan structures such as terminal sialic acid, N-acetylglucosamine and/or galactose.Therefore, a stable platform for the production and purification of recombinant hRSPO1 from HEK293 cells was generated, leading to the production of a purified, fully characterized and biologically active protein product to be applied in Tissue Engineering.
Gabriel Levin; Bruna Koga; Gustavo Belchior; Ana Carreira; Mari Sogayar

Methylome and transcriptome analyses reveal insights into the epigenetic basis for the good survival of hypomethylated ER-positive breast cancer subtype.

Journal: Clinical epigeneticsJanuar/21/2020
Breast cancer (BRCA) is a heterogeneous disease, characterized by different histopathological and clinical features and responses to various therapeutic measures. Despite the research progress of DNA methylation in classification and diagnosis of BRCA and the close relationship between DNA methylation and hormone receptor status, especially estrogen receptor (ER), the epigenetic mechanisms in various BRCA subtypes and the biomarkers associated with diagnostic characteristics of patients under specific hormone receptor status remain elusive.In this study, we collected and analyzed methylation data from 785 invasive BRCA and 98 normal breast tissue samples from The Cancer Genome Atlas (TCGA) database. Consensus classification analysis revealed that ER-positive BRCA samples were constitutive of two distinct methylation subgroups; with the hypomethylated subgroup showing good survival probability. This finding was further supported by another cohort of ER-positive BRCA containing 30 subjects. Additionally, we identified 977 hypomethylated CpG loci showing significant associations with good survival probability in ER-positive BRCA. Genes with these loci were enriched in cancer-related pathways (e.g., Wnt signaling pathway). Among them, the upregulated 47 genes were also in line with good survival probability of ER-positive BRCA, while they showed significantly negative correlations between their expression and methylation level of certain hypomethylated loci. Functional assay in numerous literatures provided further evidences supporting that some of the loci have close links with the modulation of tumor-suppressive mechanisms via regulation gene transcription (e.g., SFRP1 and WIF1).Our study identified a hypomethylated ER-positive BRCA subtype. Notably, this subgroup presented the best survival probability compared with the hypermethylated ER-positive and hypomethylated ER-negative BRCA subtypes. Specifically, we found that certain upregulated genes (e.g., SFRP1 and WIF1) have great potential to suppress the progression of ER-positive BRCA, concurrently exist negative correlations between their expression and methylation of corresponding hypomethylated CpG loci. Therefore, our study indicates that different epigenetic mechanisms likely exist in ER-positive BRCA and provides novel clinical biomarkers specific to ER-positive BRCA diagnosis and therapy.
Xiao-Qiong Chen; Fan Zhang; Qi-Chen Su; Chi Zeng; Fu-Hui Xiao; Yu Peng

All-trans-retinoid acid induces the differentiation of P19 cells into neurons involved in the PI3K/Akt/GSK3β signaling pathway.

Journal: Journal of cellular biochemistryJanuar/21/2020
The pluripotent mouse embryonal carcinoma cell line P19 is widely used as a model for research on all-trans-retinoid acid (RA)-induced neuronal differentiation; however, the signaling pathways involved in this process remain unclear. This study aimed to reveal the molecular mechanism underlying the RA-induced neuronal differentiation of P19 cells. Real-time quantitative polymerase chain reaction and Western blot analysis were used to determine the expression of neuronal-specific markers, whereas flow cytometry was used to analyze cell cycle and cell apoptosis. The expression profiles of messenger RNAs (mRNAs) in RA-induced neuronal differentiation of P19 cells were analyzed using high-throughput sequencing, and the functions of differentially expressed mRNAs (DEMs) were determined by bioinformatics analysis. RA induced an increase in both class III β-tubulin (TUBB3) and neurofilament medium (NEFM) mRNA expression, indicating that RA successfully induces neuronal differentiation of P19 cells. Cell apoptosis was not affected; however, cell proliferation decreased. We found 4117 DEMs, which were enriched in the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway, Wnt signaling pathway, and cell cycle. Particularly, a few DEMs could be identified in the PI3K/Akt signaling pathway networks, such as PI3K, Akt, glycogen synthase kinase-3β (GSK3β), cyclin-dependent kinase 4 (CDK4), P21, and Bax. RA significantly increased the protein expression of PI3K, Akt, phosphorylated Akt, GSK3β, phosphorylated GSK3β, CDK4, and P21, but it reduced Bax protein expression. The Akt inhibitor affected the increase of TUBB3 and NEFM mRNA expression in RA-induced P19 cells. The molecular mechanism underlying the RA-induced neuronal differentiation of P19 cells is potentially involved in the PI3K/Akt/GSK3β signaling pathway. The decreased cell proliferation ability of neuronally differentiated P19 cells could be associated with the expression of cell cycle proteins.
Fang Fu; Lu-Shan Li; Ru Li; Qiong Deng; Qiu-Xia Yu; Xin Yang

Identification and characterization of differentially expressed lncRNA in 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced cleft palate.

Journal: Human & experimental toxicologyJanuar/21/2020
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous environmental pollutant and also a strong teratogen for cleft palate (CP). But up to now, the underlying molecular mechanisms of TCDD-induced CP are largely unknown. More recently, accumulating evidences are revealing important roles of long noncoding RNAs (lncRNAs) in all kinds of diseases including CP. However, the role and molecular mechanism of lncRNAs in TCDD-induced CP are still largely unexplored. Thus, identification of differentially expressed lncRNA (DEL) might help figuring out the mechanism of CP induced by TCDD. In this study, a CP offspring model of C57BL/6 female mice was generated by TCDD (64 µg/kg body weight) induce on embryo day 10 (E10). The incidence rate of CP was 100% in the TCDD group (105) after cervical dislocation on E16. Then, the high-throughput RNA sequencing (RNA-seq) was established to search a comprehensive profile of the lncRNAs. In addition, a coexpression network of lncRNA and messenger RNA (mRNA) was performed to discern potential mechanism. The result showed that 26,246 novel lncRNAs and 9635 known lncRNAs were screened out, and 413 lncRNA transcripts and 65 mRNA transcripts were identified as being significantly different between the CP group and control group. Notably, we found that there are seven lncRNAs that can target Smad1 and Smad5, which are key molecules of bone morphogenetic protein (BMP) signaling pathway, which suggested that they may be concerned with BMP signaling in TCDD-induced CP. In addition, some lncRNAs targeted the important molecules of Hippo and Wnt signaling pathways. These results suggested that characteristic lncRNA alterations may play a critical role in TCDD-induced CP, which provided a theoretical basis for further research.
L-Y Gao; X-L Hao; L Zhang; T Wan; J-Y Liu; J Cao

Thyroid Hormone Actions and Bone Remodeling – The Role of the Wnt Signaling Pathway.

Journal: Experimental and clinical endocrinology & diabetes : official journal, German Society of Endocrinology [and] German Diabetes AssociationJanuar/20/2020
Thyroid hormones are indispensable for bone development and growth. Also in adults, bone mass maintenance is under the control of thyroid hormones. Preclinical and clinical studies established untreated hyperthyroidism as a cause for secondary osteoporosis with increased fracture risk. Thus, normal thyroid function is essential for bone health. Mechanistically, thyroid hormone excess accelerates bone turnover with predominant bone resorption. How thyroid hormones affect osteoblast and osteoclast functions, however, still remains ill-defined. The Wnt signaling pathway is a major determinant of bone mass and strength as it promotes osteoblastogenesis and bone formation, while inhibiting bone resorption. So far, only few studies investigated a possible link between thyroid hormones, bone metabolism and the Wnt pathway. In this review, we summarize the literature linking thyroid hormones to bone homeostasis through Wnt signaling and discuss its potential as a therapeutic approach to treat hyperthyroidism-induced bone loss.
Franziska Lademann; Elena Tsourdi; Lorenz Hofbauer; Martina Rauner

Proteome alterations in aqueous humour of primary open angle glaucoma patients.
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Journal: International journal of ophthalmologyJanuar/20/2020
To unravel the primary open angle glaucoma (POAG) related proteomic changes in aqueous humour (AH).
Totally 35 patients listed for cataract surgery (controls: n=12, age: 67.4±13.6y) or trabeculectomy for POAG (n=23, age: 72.5±8.3y) were included. AH samples of those patients were obtained during cataract surgery or trabeculectomy. AH samples were subsequently pooled into the experimental groups under equal contribution in terms of protein amount of each individual patient. Protein samples were analyzed by a linear trap quadrupol Orbitrap Mass Spectrometry device with an upstream liquid chromatography system. The obtained raw data were analyzed using the Maxquant proteome software and compared. Proteins with a fold-change ratio higher than a cut-off of 2 were considered as noticeably altered.
A total number of 175 proteins could be identified out of the AH from POAG and cataract by means of quantitative mass spectrometric analysis. Apolipoprotein D (fold change, 3.16 times), complement C3 (2.96), pigment epithelium-derived factor (2.86), dickkopf-related protein 3 (2.18) and wingless-related integration (Wnt) inhibitory factor 1 (2.35) were significantly upregulated within the AH of glaucoma compared to cataract serving as controls.AH provides a tool to analyze changes in glaucoma and shows striking changes in Wnt signaling inhibitory molecules and other proteins.
Hanhan Liu; Fabian Anders; Sebastian Funke; Karl Mercieca; Franz Grus; Verena Prokosch

Effects of propofol on colon cancer metastasis through STAT3/HOTAIR axis by activating WIF-1 and suppressing Wnt pathway.

Journal: Cancer medicineJanuar/18/2020
In the present study, we aim to investigate the potential role of propofol in the tumor progression of colon cancer.Human colon cancer cell lines were cultured and exposed with 8 μg/mL propofol. RNA interference was performed to silence the expression of HOTAIR or STAT3 to explore their biological functions in colon cancer. Cell apoptosis and invasion were assessed using flow cytometry and transwell assays, respectively. Quantitative real-time PCR, western blot, and immunohistochemistry were subjected to measure the expression patterns of HOTAIR, STAT3, Wnt signaling factors, and epithelial-mesenchymal transition-related markers, respectively. Besides, nude mice were transplanted with colon cancer cells for further exploration. Tumor formation, volume, and weight were evaluated to validate the in vitro findings.Propofol treatment promoted cell apoptosis and inhibited cell invasion in colon cancer cells, while the effects were reversed by HOTAIR overexpression. Additionally, STAT3 positively regulated HOTAIR expression, which was also negatively modulated by propofol. Moreover, STAT3 and HOTAIR were shown to independently regulate colon cancer cell apoptosis and invasion. Furthermore, HOTAIR could stimulate Wnt signaling pathway via inhibiting WIF-1 expression and upregulating β-catenin expression, which was also demonstrated by in vivo study.Taken together, the current study demonstrated that propofol exerts the inhibition on cell invasion and promotion on cell apoptosis through regulating STAT3/HOTAIR by activating WIF-1 and suppressing Wnt pathway, indicating that propofol might serve as a therapeutic role for colon cancer patients in the future.
Yun-Fei Zhang; Chang-Sheng Li; Yi Zhou; Xi-Hua Lu

Targeting the Wnt signalling pathway: the challenge of reducing scarring without affecting repair.

Journal: Expert opinion on investigational drugsJanuar/17/2020
Introduction: Globally, deaths from liver disease are increasing and for most patients there are few curative options. Fibrosis or scarring is often associated with the formation and progression of liver disease; however, clinical anti-fibrotic therapies are lacking. Recent work has shown that Wnt signalling, a signalling pathway that is necessary for embryonic development and cancer, can also regulate scar formation in the liver.Areas covered: This article seeks to shed light on the dualistic role of Wnt signalling in liver regeneration following injury and how Wnt signalling can regulate scar formation. It also discusses how Wnt signalling cooperates with other classical fibrogenic signalling cascades, such as TGFβ signalling. Finally, the article examines recent advances in the development of Wnt signalling pathway inhibitors and asks whether repurposing these agents as anti-fibrotic therapies is a realistic option.Expert Opinion: The understanding of Wnt signalling in liver regeneration and fibrosis is in its infancy and whilst new generations of Wnt pathway inhibitors have shown anti-fibrotic effects, further research is necessary to enhance our understanding of the Wnt-landscape in different patterns of liver disease. This will accelerate the development of more specific Wnt inhibitor-based anti-fibrotics.
Edward Jarman; Luke Boulter

Correction to: ZNF326 promotes malignant phenotype of glioma by up-regulating HDAC7 expression and activating Wnt pathway.

Journal: Journal of experimental & clinical cancer research : CRJanuar/16/2020
In the original publication of this manuscript [1], the author mislabeled the CTL group and ZNF326 group in Fig. 2-I,J (MTT result). The revised Fig. 2 is shown below.
Xinmiao Yu; Minghao Wang; Jingjing Wu; Qiang Han; Xiupeng Zhang

Expression of Wnt/β-catenin related genes after skeletal muscle contusion.

Journal: International journal of clinical and experimental pathologyJanuar/15/2020
It was aimed to determine expressions of genes related to Wnt/β-catenin signaling for evaluating time duration after skeletal muscle contusion.Pathological change of skeletal muscle was observed after H-E staining. mRNA of respective genes was quantified with real-time quantitative PCR. Expression of β-catenin was further characterized with immunostaining and quantified as intensity/area and further immune blotting and quantified as grey intensity normalized to loading control (GADPH).After injury, skeletal muscle exhibited prominent inflammatory response, hyperplasia and regeneration. Infiltration of inflammatory cell, formation of myotube and maturation of skeletal muscle fiber were observed under HE staining. Expression of FZD4, Myo D, Myf5 changed during early stages after injury and could serve to evaluate injury within 24 h; Expression of SFRP5 and Fra1 changed during early-to-intermediate stages after injury and could serve to evaluate injury within 12-48 h; Expression of MRF4 changed during intermediate stages after injury and could serve to evaluate injury within 36-48 h; Expression of β-catenin changed during intermediate stages after injury and could serve to evaluate injury within 36 h-3 d; Expression of MyoG changed during late stages after injury and could serve to evaluate injury within 48 h-7 d. Immunostaining experiments showed that 36 h after injury, membrane β-catenin decreased while nucleus β-catenin increased.Wnt/β-catenin related genes are involved in regeneration of skeletal muscle after contusion. The sequential changes of gene expression can be used for evaluating the duration after contusion.
Xin-Hua Liang; Zhi-Jie Liu; Jun-Hong Sun; Zu-Xin Dong; Jian Lu; Mei-Ling Jiang+2 authors

Aberrant Expressional Profiling of Known MicroRNAs in the Liver of Silver Carp (Hypophthalmichthys molitrix) Following Microcystin-LR Exposure Based on samllRNA Sequencing.

Journal: ToxinsJanuar/15/2020
Microcystin-LR (MC-LR) poses a serious threat to human health due to its hepatotoxicity. However, the specific molecular mechanism of miRNAs in MC-LR-induced liver injury has not been determined. The aim of the present study was to determine whether miRNAs are regulated in MC-LR-induced liver toxicity by using high-throughput sequencing. Our research demonstrated that 53 miRNAs and 319 miRNAs were significantly changed after 24 h of treatment with MC-LR (50 and 200 μg/kg, respectively) compared with the control group. GO enrichment analysis revealed that these target genes were related to cellular, metabolic, and single-organism processes. Furthermore, KEGG pathway analysis demonstrated that the target genes of differentially expressed miRNAs in fish liver were primarily involved in the insulin signaling pathway, PPAR signaling pathway, Wnt signaling pathway, and transcriptional misregulation in cancer. Moreover, we hypothesized that 4 miRNAs (miR-16, miR-181a-3p, miR-451, and miR-223) might also participate in MC-LR-induced toxicity in multiple organs of the fish and play regulatory roles according to the qPCR analysis results. Taken together, our results may help to elucidate the biological function of miRNAs in MC-LR-induced toxicity.
Yiyi Feng; Xi Chen; Junguo; Bangjun Zhang; Xiaoyu Li

Exosomes for Diagnosis and Therapy in Gastrointestinal Cancers.

Journal: International journal of molecular sciencesJanuar/15/2020
Exosomes are membrane-bound extracellular vesicles (EVs) released by most cells, having a size ranging from 30 to 150 nm, and are involved in mechanisms of cell-cell communication in physiological and pathological tissues. Exosomes are engaged in the transport of biomolecules, such as lipids, proteins, messenger RNAs, and microRNA, and in signal transmission through the intercellular transfer of components. In the context of proteins and nucleic acids transported from exosomes, our interest is focused on the Frizzled proteins family and related messenger RNA. Exosomes can regenerate stem cell phenotypes and convert them into cancer stem cells by regulating the Wnt pathway receptor family, namely Frizzled proteins. In particular, for gastrointestinal cancers, the Frizzled protein involved in those mechanisms is Frizzled-10 (FZD-10). Currently, increasing attention is being devoted to the protein and lipid composition of exosomes interior and membranes, representing profound knowledge of specific exosomes composition fundamental for their application as new delivering drug tools for cancer therapy. This review intends to cover the most recent literature on the use of exosome vesicles for early diagnosis, follow-up, and the use of these physiological nanovectors as drug delivery systems for gastrointestinal cancer therapy.
Maria Scavo; Nicoletta Depalo; Valeria Tutino; Valentina De Nunzio; Chiara Ingrosso; Federica Rizzi+3 authors

Effects of melatonin on diabetic nephropathy rats via Wnt/β-catenin signaling pathway and TGF-β-Smad signaling pathway.

Journal: International journal of clinical and experimental pathologyJanuar/15/2020
This research aimed to explore the protective effect of melatonin on diabetic nephropathy (DN) rats induced by streptozotocin (STZ) and its related signaling pathways.100 SPF male Sprague-Dawley rats were divided into four groups: low dose melatonin group, medium dose melatonin group and high dose melatonin group. Rats were 35 mg/kg STZ once to establish a DN model, and control rats were given the corresponding dose of normal saline. A renal function test was used to measure urine protein (UP), blood urea nitrogen (BUN) and serum creatinine (Scr). Pathological changes of renal tissues were obtained by HE staining and Masson staining. Oxidative stress-related indicators were measured in a STZ-induced DN rat. Western blot was used to measure target proteins in renal tissues.
The levels of UP, BUN and Scr in the model group were significantly higher than control group (P<0.05). After administration of melatonin, each administration group was significantly decreased compared to the model group. Pathological changes of renal tissues in the high dose group were the closest to the control group. After administration of melatonin, activities of SOD, CAT and GSH-Px were significantly increased in the medium dose group and the high dose group (P<0.05), while the activity of MDA was significantly decreased (P<0.05). The expression of Wnt4 and β-catenin in the model group were higher than the control group (P<0.01). When melatonin was given, the expression of Wnt4 and β-catenin in the medium dose group and the high dose group were significantly lower than the model group. Levels of TGF-β1, p-Samd2 and p-Samd3 in the control group were lower than the model group (P<0.05), and were decreased in the medium dose group and the high dose group.
Melatonin improves renal function, relieves oxidative stress, and protects the renal tissue via the Wnt/β-catenin signaling pathway and the TGF-β1-Smad2/3 signaling pathway in STZ-induced DN rats.
Weichao Wang; Jie Zhang; Xiaohang Wang; Hong Wang; Qiaohua Ren; Yukun Li

Combined effects of fibroblast growth factor 2 and dexamethasone on differentiation of human cementoblasts.

Journal: International journal of clinical and experimental pathologyJanuar/15/2020
Fibroblast growth factor 2 (FGF-2) is known to play a pivotal role in bone remodeling and osteogenesis. Dexamethasone is a potent modulator of osteogenic differentiation and can increase the bone formation capacity of cementoblasts. However, the combined effects of FGF-2 and dexamethasone on cementoblasts and the way they regulate the activities of cementoblasts have not been fully understood. The aim of this study was to clarify and investigate the effects of the combination of FGF-2 and dexamethasone on growth and differentiation in human cementoblasts and to determine the underlying mechanism. A series of experiments including MTT, alkaline phosphatase (ALP) activity, alizarin red staining, RT-PCR, and Western blot were carried out to evaluate the proliferation and differentiation of cementoblasts. In addition, the changes of Wnt signaling pathway molecules were checked to analyze the possible mechanism. Compared to the treatment of cementoblasts with either FGF 2 or dexamethasone alone, the combination of FGF-2 and dexamethasone synergistically increased cell proliferation, ALP activity, nodule formation, and expression of differentiation markers. In addition, FGF 2 and dexamethasone enhanced expression of Wnt3a, Runx2, β-catenin, and p-GSK-3β, all of which were inhibited by DKK1. Taken together, FGF-2 and dexamethasone act synergistically, enhancing each other’s ability to stimulate cementoblast growth and differentiation via Wnt signaling pathways. These results support the therapeutic potential of a combination strategy for facilitating cementogenesis and aiding periodontal regeneration.
Yufei Xie; Gang Shen

Circulating microRNAs, miR-10b-5p, miR-328-3p, miR-100 and let-7, are associated with osteoblast differentiation in osteoporosis.

Journal: International journal of clinical and experimental pathologyJanuar/15/2020
Osteoporosis has become a major disease that threatened post-menopausal women and elder people. Circulating micorRNAs (miRNA) could provide useful information for diagnosis and therapeutics. The study employed RT-real time PCR to detect the circulating miRNAs between osteoporotic patients and healthy controls. Human and mouse osteoblast cell lines were used to test the differential induction effects by miRNAs. Alkaline phosphatase activity and Alizarin red staining were examined after miRNA mimics stimulation. The authors found 14 of 150 tested miRNAs were significantly aberrant expressed between patients and healthy controls. Results showed miR-328-3p, let-7g-5p, miR-133b, miR-22-3p, miR-2861, miR-518 miR-100 were down-regulated osteoporotic patient, while miR-10b-5p, miR-21, miR-125b and miR-127 were up-regulated. MiR-10b-3p, miR-328-3p, miR-100 and let-7 showed tight association with Wnt pathway. MiR-10b-5p increased ALP activity and mineral deposition in human and mouse osteoblast cells, indicating miR-10b-3p promoted osteoblast cell differentiation. MiR-328-3p and let-7g-5p decreased ALP activity and suppressed mineral deposition in both cell lines. Conclusively, miR-10b-5p promoted osteoblast cells differentiation; miR-328-3p, miR-100 and let-7 inhibited osteoblast cells differentiation.
Ruisong Chen; Xin Liao; Fengrong Chen; Bowen Wang; Jianming Huang; Guojian Jian

Diagnostic Value Investigation and Bioinformatics Analysis of miR-31 in Patients with Lymph Node Metastasis of Colorectal Cancer.
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Journal: Analytical cellular pathology (Amsterdam)Januar/14/2020
Colorectal cancer (CRC) is one of the most frequent cancers occurring in developed countries. Distant CRC metastasis causes more than 90% of CRC-associated mortality. MicroRNAs (miRNAs) play a key role in regulating tumor metastasis and could be potential diagnostic biomarkers in CRC patients. This study is aimed at identifying miRNAs that can be used as diagnostic biomarkers for CRC metastasis. Towards this goal, we compared the expression of five miRNAs commonly associated with metastasis (i.e., miR-10b, miR-200c, miR-155, miR-21, and miR-31) between primary CRC (pCRC) tissues and corresponding metastatic lymph nodes (mCRC). Further, bioinformatics analysis of miR-31 was performed to predict target genes and related signaling pathways. Results showed that miR-31, miR-21, miR-10b, and miR-155 expression was increased to different extents, while miR-200c expression was lower in mCRC than that in pCRC. Moreover, we found that the level of both miR-31 and miR-21 was notably increased in pCRC when lymph node metastasis (LNM) was present, and the increase of miR-31 expression was more profound. Hence, upregulated miR-31 and miR-21 expression might be a miRNA signature in CRC metastasis. Moreover, we detected a higher miR-31 level in the plasma of CRC patients with LNM compared to patients without LNM or healthy individuals. With the bioinformatics analysis of miR-31, 121 putative target genes and transition of mitotic cell cycle and Wnt signaling pathway were identified to possibly play a role in CRC progression. We next identified seven hub genes via module analysis; of these, TNS1 was most likely to be the target of miR-31 and had significant prognostic value for CRC patients. In conclusion, miR-31 is significantly increased in the cancer tissues and plasma of CRC patients with LNM; thus, a high level of miR-31 in the plasma is a potential biomarker for the diagnosis of LNM of CRC.
Wu-Wen Zhang; Xin-Liang Ming; Yuan Rong; Chao-Qun Huang; Hong Weng; Hao Chen+2 authors

The clinicopathological significance and prognostic value of β-catenin Ser45-phosphorylation expression in esophageal squamous cell carcinoma.

Journal: International journal of clinical and experimental pathologyJanuar/14/2020
β-Catenin is a multifunctional protein which plays a central role in physiological homeostasis, and it acts both as an adaptor protein for intracellular adhesion and a transcriptional co-regulator. As a pivotal component of the Wnt signaling pathway, the accumulation of β-catenin in the cytoplasm/nucleus leads to many diseases including cancer. The phosphorylation at Ser45 of β-catenin causes the degradation of β-catenin, which makes β-catenin at a very low level. It has been shown that phosphorylation at Ser45 of β-catenin is closely related to the occurrence and development of tumors. However, little is known about the exact role of β-catenin Ser45-phosphorylation in esophageal squamous cell carcinoma (ESCC).The present study was aimed at exploring the role and the prognostic value of the expression of β-catenin Ser45-phosphorylation in ESCC.The expression of phosphorylation at Ser45of β-catenin was detected by immunohistochemistry in 90 cases of ESCC and their corresponding adjacent nonneoplastic esophageal tissues (n = 90). We then evaluated the relationships among the expressions of phosphorylation at Ser45 of β-catenin, the clinicopathological parameters, and the prognoses of the ESCC patients.
The expression level of phosphorylation at Ser45 of β-catenin in ESCC was 65.6% (59/90), significantly lower than the expression level in nonneoplastic esophageal tissues, where it was 88.9% (80/90), (X2 = 10.340, P = 0.003). The expression of β-catenin Ser45-phosphorylation was significantly related to the degree of tumor cell differentiation, but not to age, gender, tumor size, AJCC clinical stage, or lymphatic metastasis. In univariate and multivariate Cox regression analyses, we found that lymphatic invasion and depth of invasion were independent risk factors for the poor prognosis of ESCC patients. Furthermore, a survival analysis revealed that the positive expression of β-catenin Ser45-phosphorylation had a significantly better survival date than the negative group after curative surgery.
β-catenin Ser45-phosphorylation may play an important role in the pathogenesis and development of ESCC and may provide clinically useful prognostic information.
Xinning Pan; Lili; Jinsheng Wang

MicroRNA-23 inhibition protects the ischemia/reperfusion injury via inducing the differentiation of bone marrow mesenchymal stem cells into cardiomyocytes.

Journal: International journal of clinical and experimental pathologyJanuar/14/2020
Recently, miRNA-23 has been illustrated to play an important role in causing myocardial ischemia/reperfusion injury (MIRI), indicated that inhibition of miR-23 could protect the cardiomyocyte from MIRI. However, the underlying mechanism of miR-23 inhibition in alleviating the reperfusion-induced myocardial damage is unclear. Recognizing that the bone marrow mesenchymal stem cells (BMSCs) have the potential for pluripotent differentiation into myocardial cells, we therefore hypothesis that the BMSCs are involved in the process of miR-23 alleviating IRI. For verification, the BMSCs was established firstly and confirmed by the immunofluorescence assay and flow cytometry analysis. As results revealed that BMSCs were positive for CD44 which was known for BMSC markers, and negative expression for CD45, indicating that the BMSCs was successfully established in our work. Subsequently, we have investigated the effect of miR-23 on the expression of hyaluronan synthase-2 (Has2), a critical gene during heart morphogenesis. Results obtained by the Western-blot and qRT-PCR assay displayed that the levels of Has2 in the BMSCs treated by miR-23 inhibitor was significantly up-regulated than that of control group. Furthermore, the effect of miR-23 on promoting the transformation of BMSCs into myocardial cells was investigated. As demonstrated by the results that the expression level of the cardiac markers in BMSCs transfected with miR-23 inhibitor was remarkably elevated, indicating that inhibition of miR-23 exactly facilitated to the transformation of BMSCs into myocardial cells. The underlying mechanisms experiments showed that the Wnt1, TCF4, and the β-catenin could be significantly elevated by treating with miR-23 inhibitor, suggesting that the activation of Wnt pathway has played a significant role in that process. Finally, the in vivo IRI antagonism effect of miR-23 inhibition was studied and results displayed that the myocardium lesions of these IR rats could be significantly recovered by treating with miR-23 inhibitor.
Mingjun Lu; Yongzhe Xu; Min Wang; Tao Guo; Fuquan Luo; Nan Su+3 authors

Wnt/β-catenin signaling regulates pathogenesis of human middle ear cholesteatoma.

Journal: International journal of clinical and experimental pathologyJanuar/14/2020
Cholesteatoma is characterized by the presence of a squamous epithelium invading the middle ear altering its growth properties. Wnt/β-catenin signaling controls cell proliferation and differentiation by regulating expressions of target genes. Elevated levels of β-catenin are related to tissue pathogenesis and tumor progression. Nevertheless, the mechanisms through which β-catenin contributes to middle ear cholesteatoma development remain to be elucidated. We used proliferation assay, qRT-PCR assay, and western blotting to measure levels of the Wnt/β-catenin signaling pathway. β-Catenin expression evidently increased in middle ear cholesteatoma cells when compared with normal epithelial cells. Next, we found that treatment of Wnt inhibitor dickkopf1 (Dkk1) decreased β-catenin expression, as well as the expression levels of cytokeratin 16 (CK16), CK18, Ki67 and PCNA. Overexpression of Wnt3a or β-catenin induced the expression levels of CK16, CK18, Ki67 and PCNA. Furthermore, Dkk1 treatment significantly inhibited proliferation activity of middle ear cholesteatoma cells, whereas forced expression of Wnt3a or β-catenin promoted proliferation activity of middle ear cholesteatoma cells. Wnt/β-catenin signaling induced cell proliferation and up-regulated expressions of targeted genes in human middle ear cholesteatoma.
Juan Lin; Qing Ye; Yihong Wang; Ying Wang; Yanfen Zeng

miR-601 inhibits proliferation, migration and invasion of prostate cancer stem cells by targeting KRT5 to inactivate the Wnt signaling pathway.

Journal: International journal of clinical and experimental pathologyJanuar/14/2020
This study aimed to verify the hypothesis that downregulation of miR-601 inhibits the proliferation, migration, and invasion of prostate cancer stem cells (PCSCs) by the Wnt signaling pathway through targeting keratin 5 (KRT5).Bioinformatic tools were applied to predict miRNAs and genes potentially associated with prostate cancer, then miR-601 and KRT5 were selected. Subsequently, PCSCs were investigated with respect to miR-601 overexpression or inhibition, KRT5 overexpression, or treatment with a Wnt pathway inhibitor. A series of experiments including western blotting, RT-qPCR, wound healing experiment, transwell assay, MTT assay, annexin V-FITC/PI flow cytometric analysis, nude mice assay and immunohistochemistry were then carried out.
Compared with negative control group, migration, invasion, and proliferation of PCSCs and Wnt-1 expression were all enhanced, but apoptosis was attenuated in the miR-601 mimic group. Furthermore, results identified in the other groups (KRT5, miR-601 inhibitor, miR-601 inhibitor + KRT5, Wnt signaling pathway inhibitor, PRI-724/PRI-724 + KRT5) were opposite to those identified with the miR-601 mimic group (all P<0.05). Compared with the miR-601 inhibitor + KRT5 group, migration, invasion, and proliferation of PCSCs and Wnt-1 expression were all increased, whereas apoptosis was suppressed in KRT5 or miR-601 inhibitor groups (all P<0.05). Compared with the PRI-724 + KRT5 group, migration, invasion, and proliferation of PCSCs and Wnt-1 expression were also enhanced, whereas apoptosis was inhibited in PRI-724 or KRT5 groups (all P<0.05).
Results obtained from the present study have demonstrated that downregulation of miR-601 is able to inhibit the proliferation, migration, and invasion of PCSCs by activating KRT5, and subsequently inhibiting the Wnt pathway.
Hongbing Du; Xinghuan Wang; Rui Dong; Dongliang Hu; Yaoyi Xiong

Expression of ARHGAP10 correlates with prognosis of prostate cancer.

Journal: International journal of clinical and experimental pathologyJanuar/14/2020
Prostate cancer is one of the most common malignancies in men worldwide. Altered expression of ARHGAP10, a member of the Rho GTPase activating protein (RhoGAP) family, has been found in several human cancers. However, its clinical significance in prostate cancer remains unknown. In the current study, we found that mRNA levels of ARHGAP10 were significantly higher in prostate cancer tissues than in the non-cancerous controls. Gene set enrichment analysis (GSEA) revealed that ARHGAP10 expression was negatively correlated with the Wnt signaling pathway. Immunohistochemical staining results showed that 62.2% (56/90) and 65.5% (59/90) of prostate cancer tissues displayed low expression of ARHGAP10 and high expression of β-catenin, respectively. ARHGAP10 protein expression was significantly correlated with histologic grade (P < 0.0001), tumor stage (P = 0.0298), preoperative prostate specific antigen level (P = 0.0261), vital status (P = 0.0017) and β-catenin expression (P < 0.0001). Kaplan-Meier survival analysis indicated that patients with low levels of ARHGAP10 and high levels of β-catenin had poor overall survival. Multivariate analyses revealed that ARHGAP10 and β-catenin expression was independent prognostic factor for prostate cancer. In summary, the current study suggests that ARHGAP10 in association with β-catenin may play a role in the development of prostate cancer and serve as a prognostic factor for this disease.
Hua Gong; Xingyi Chen; Yongcao Jin; Jiasun Lu; Yuanjue Cai; Ouyang Wei+5 authors

Study of the Gastrointestinal Heat Retention Syndrome in Children: From Diagnostic Model to Biological Basis.
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Journal: Evidence-based complementary and alternative medicine : eCAMJanuar/13/2020
Gastrointestinal heat retention syndrome (GHRS) refers to a condition that is associated with increased gastrointestinal heat caused by a metabolic block in energy. It is common in children and is closely related to the occurrence and development of recurrent respiratory tract infection, pneumonia, recurrent functional abdominal pain, etc. However, there are no standardized diagnostic criteria to differentiate the GHRS. Therefore, this study is aimed to establish a diagnostic model for children’s GHRS and explore the possible biological basis by using systems biology to achieve. Furthermore, Delphi method and the clinical data of Lasso analysis were used to screen out the core symptoms. Nineteen core symptoms of GHRS in children were screened including digestive symptoms such as dry stool, poor appetite, vomiting, and some nervous system symptoms such as night restlessness and irritability. Based on the core symptoms, a GHRS diagnosis model was established using the eXtreme Gradient Boosting (XGBoost) method, and the accuracy of internal verification reached 93.03%. Relevant targets of the core symptoms in the Human Phenotype Ontology (HPO) were retrieved, and target interactions were linked through the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database, and core targets were selected after topological analysis using Cytoscape. Relevant biological processes and pathways were analyzed by applying the DAVID and KEGG databases. The enriched biological processes focused on the cell proliferation, differentiation, apoptosis, and mitochondrial metabolism, which were mainly associated with PI3K-AKT, MAPK network pathways, and the Wnt signaling pathway. In conclusion, we established a diagnosis model of GHRS in children based on the core symptoms and provided an objective standard for its clinical diagnosis. And, the Wnt signaling pathway and the estrogen receptor-activated PI3K-AKT and MAPK network pathways may play important roles in the GHRS processing.
Xueyan; Chencheng Mei; Ling Huang; Chen Bai; Jingnan Xu; Yuxiang Wan+7 authors

Prognostic Impact of Lymphoid Enhancer Factor 1 Expression and Serum Galectin.3 in Egyptian AML Patients.
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Journal: Advances in hematologyJanuar/13/2020
Deregulation of the Wnt signaling pathway had a role in haematological malignancies. Previous studies reported that lymphoid enhancer factor 1 (LEF1) expression and serum Galectin-3 level could affect clinical parameters and outcome in acute myeloid leukemia patients, but as far as we know, no study has addressed their combined effect on AML patients.We studied the expression of LEF1 by real-time qPCR and measured serum level of Gal.3 by ELISA technique in peripheral blood of 69 AML patients and correlated it with different clinicopathological criteria of patients, response, PFS and OS.
We found high expression (LEF1high) was associated with better OS (p = 0.02) and EFS (p = 0.019) compared to LEF1low, low serum Gal.3 level had better OS (p = 0.014) and EFS (p = 0.02) compared to high serum Gal.3 level. LEF1high less likely to carry a FLT3-ITD (p = 0.047) compared to LEF1low patient, also LEF1high characterized by favorable risk (p = 0.02) than LEF1low patients. While patients with higher Gal-3 levels characterized by poor risk (p = 0.02) than lower Gal.3 lels, also more likely to carry a FLT3-ITD with borderline significance (p = 0.054). Combined LEF1high/Gal.3 low patients had lower baseline blast percentages (p = 0.02), favorable risk (p = 0.01), less likely to carry FLT3-ITD (p = 0.02), higher CR rate (p = 0.055), shorter time to CR (0.001) than other groups. Among high Gal.3 level group, LEF1highexpression improved OS and EFS (20 and 15 months respectively) vs LEF1low expression (13 and 8 months respectively).
We conclude that high LEF1 expression was a favorable prognostic marker which can define AML patient risk and outcome independent from assessing the serum galectin.3 level.
M ElBaiomy; S Aref; M Zaafarany; Sara Atwa; Tamer Akl; Wafaa El-Beshbishi+3 authors

Attenuation of Wnt/β-catenin signaling in patients with Stevens-Johnson syndrome and toxic epidermal necrolysis.
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Journal: International journal of biological sciencesJanuar/13/2020
Stevens-Johnson syndrome (SJS) and toxic epidermal necrosis (TEN) are rare but life-threatening severe cutaneous adverse reactions. Current studies have suggested that the pathobiology of drug-mediated SJS/TEN involves a dysregulation of cellular immunity with overwhelming activation of cytotoxic T lymphocytes. The canonical Wnt signaling pathway plays important roles in T cell development and activation, which may provide potential avenues for alleviating dysregulated immunity in SJS/TEN. In this study, we aimed to assess the implication of Wnt signaling in drug-reactive T cells in SJS/TEN. We showed downregulation of Wnt signaling components, including T cell factor 1 (TCF-1)/lymphoid enhancer binding factor 1 (LEF-1) transcription factors, in SJS/TEN patients, suggesting that canonical Wnt signaling is regulated during cytotoxic T cell responses in SJS/TEN. Further analyses demonstrated that engagement of the T cell receptor by antigen encounter and treatment of a prognostic marker of SJS/TEN, IL-15, in vitro led to the downregulation of LEF-1 and TCF-1 expression in CD8+ T cells. Enhancement of Wnt signaling by adding the Wnt activators attenuated ex vivo activation of drug-specific T cells from SJS/TEN patients, indicating a functional involvement of Wnt signaling in the pathomechanism of SJS/TEN. These findings provide additional insight into the immunopathogenesis and therapeutic intervention of this devastating condition.