DANCR Regulates hESC Differentiation Towards Definitive Endoderm / 中山大学学报(医学科学版)

ObjectiveTo explore the function of DANCR during the differentiation of human embryonic stem cells （hESC） toward definitive endoderm （DE）. MethodsThe in vitro DE differentiation system was established and its efficiency was verified. The correlation between the expression level of DANCR and DE differentiation process was detected. Using lentivirus system， we stably knocked down DANCR in hESC. The shDANCR hESC line was applied to DE differentiation， using qPCR and Western blot to detect the expression of DE marker genes SOX17 and FOXA2， and that of primitive streak marker genes Brachyury （T）， EOMES， MIXL1 and GSC. Dual luciferase reporter assay and qPCR were used to confirm the interaction between DANCR and the WNT pathway during DE differentiation. ResultsThe in vitro differentiation system mimicked DE differentiation efficiently. And the expression of DANCR was gradually downregulated during differentiation. DANCR was efficiently knocked down in the shDANCR hESC line （P < 0.001）. Compared with those in the control group， the expression levels of primitive markers Brachyury （T）， EOMES， MIXL1 and GSC， as well as DE markers SOX17 and FOXA2， were significantly decreased in shDANCR groups （P < 0.05）. Furthermore， the transcriptional activity of the WNT pathway in shDANCR groups was lower than that in the control group （P < 0.05）. And RNA levels of downstream genes of the WNT pathway， FZD5， FZD8， SFRP1， FRZB and ANKRD6， were significantly decreased in shDANCR groups （P < 0.05）. However， differences in protein levels of the TGFβ pathway effectors SMAD2/3 and p-SMAD2 were statistically insignificant in shDANCR and control groups （P > 0.05）. Forced activation of β-CATENIN rescued DANCR knock down-induced deficiency in DE differentiation. ConclusionsThe expression of DANCR decreases during DE differentiation. DANCR may promote DE differentiation through modulating the activity of the WNT pathway.

A novel TNKS/USP25 inhibitor blocks the Wnt pathway to overcome multi-drug resistance in TNKS-overexpressing colorectal cancer

Modulating Tankyrases (TNKS), interactions with USP25 to promote TNKS degradation, rather than inhibiting their enzymatic activities, is emerging as an alternative/specific approach to inhibit the Wnt/β-catenin pathway. Here, we identified UAT-B, a novel neoantimycin analog isolated from Streptomyces conglobatus, as a small-molecule inhibitor of TNKS-USP25 protein-protein interaction (PPI) to overcome multi-drug resistance in colorectal cancer (CRC). The disruption of TNKS-USP25 complex formation by UAT-B led to a significant decrease in TNKS levels, triggering cell apoptosis through modulation of the Wnt/β-catenin pathway. Importantly, UAT-B successfully inhibited the CRC cells growth that harbored high TNKS levels, as demonstrated in various in vitro and in vivo studies utilizing cell line-based and patient-derived xenografts, as well as APCmin/+ spontaneous CRC models. Collectively, these findings suggest that targeting the TNKS-USP25 PPI using a small-molecule inhibitor represents a compelling therapeutic strategy for CRC treatment, and UAT-B emerges as a promising candidate for further preclinical and clinical investigations.

Expression and Significance of Leucine-rich Repeat-containing G-protein Coupled Receptor 5/6 in Wnt Pathway in Children with Acute Lymphoblastic Leukemia / 中国医学科学院学报

Objective To study the expression and significance of leucine-rich repeat-containing G-protein coupled receptor(LGR)5/6 in childhood acute lymphoblastic leukemia(ALL). Methods A total of 39 children who had ALL and achieved complete remission on day 33 after induction therapy were enrolled.The children before induction therapy were considered as the incipient group,and those who achieved complete remission on day 33 by induction therapy were considered as the remission group.According to the degree of risk,they were assigned into 3 groups:low-risk(

IWR-1-endo affects the migration and proliferation of hepatocarcinoma cells by inhibiting the Wnt pathway / 国际肿瘤学杂志

Objective:To investigate the effects and mechanisms of Wnt pathway inhibitor IWR-1-endo on the biological behaviors of human hepatocarcinoma cell Huh7.Methods:Human hepatocellular carcinoma cell Huh7 was cultured in vitro, and Huh7 cells were treated with IWR-1-endo at different concentrations (0, 20, 40, 80, 160, 320 μmol/L). Scratch test was used to detect changes in cell migration ability at diffe-rent drug concentrations, plate cloning was used to detect changes in cell proliferation, Western blotting was used to detect changes in the expression of Wnt pathway related protein β-catenin, and immunofluorescence staining was used to detect the expression of β-catenin in cytoplasm and nucleus. Results:The results of the scratch test showed that the 24 h scratch healing rates of Huh7 cells treated with 0, 20, 40, 80, 160, 320 μmol/L IWR-1-endo were (20.55±0.05)%, (12.10±0.08)%, (9.36±0.10)%, (3.62±0.09)%, (0.62±0.04)% and (0.23±0.02)%, respectively, and there was a statistically significant difference ( F=230.87, P<0.001). Further pair comparison showed that there were statistically significant differences in 24 h scratch healing rates among different concentrations (all P<0.001). The 48 h scratch healing rates were (34.77±0.08)%, (17.69±0.05)%, (11.60±0.04)%, (5.68±0.07)%, (2.66±0.04)% and (1.75±0.02)%, respectively, and there was a statistically significant difference ( F=589.68, P<0.001). Further pair comparison showed that there were statistically significant differences in 48 h scratch healing rates among different concentrations (all P<0.001). After treatment with IWR-1-endo at the concentration of 0, 20, 40, 80, 160, 320 μmol/L, the clone formation rates of Huh7 cells were (61.67±0.21)%, (57.33±0.11)%, (50.00±0.25)%, (36.67±0.28)%, (23.33±0.12)% and (15.00±0.08)%, respectively, and there was a statistically significant difference ( F=403.56, P<0.001). Further pair comparison showed that there were statistically significant differences in clone formation rates among different concentrations (all P<0.001). After treatment with 0, 20, 40, 80, and 160 μmol/L IWR-1-endo for 24 h, the relative expression levels of β-catenin in Huh7 cells were 0.30±0.08, 0.25±0.07, 0.22±0.05, 0.15±0.01 and 0.06±0.02, respectively, and there was a statistically significant difference ( F=247.00, P<0.001). Compared with 0 μmol/L, the relative expression levels of β-catenin treated with 80 and 160 μmol/L had statistical significance ( P=0.014; P=0.008). Compared with 0 mol/L, immunofluorescence showed that the expressions of β-catenin in cytoplasm and nucleus were reduced after 80 μmol/L IWR-1-endo treatment. Conclusion:Wnt pathway inhibitor IWR-1-endo can inhibit the migration and proliferation of hepatocarcinoma cells Huh7 by inhibiting the activity of Wnt pathway. The above inhibitory effects are dose-dependent.

Targeting the canonical Wnt/β-catenin pathway in cancer radioresistance: Updates on the molecular mechanisms

Radiation resistance is an important factor that affects the efficacy of radiotherapy; it could even lead to its failure. In recent years, the relationship between the classical Wnt signaling pathway and radiation resistance has gradually attracted attention from scholars. Although most of the findings are comprehensive, they are fragmented and disorganized. This review explores the relationship between classical Wnt signaling pathways and cancer radiation resistance. Previous literature regarding the classical Wnt signaling pathways and cancer radiation resistance from the past decades had been summarized in this article. Moreover, the molecular mechanisms and functions of the canonical Wnt signaling pathway involved in the formation of radioresistance were systemically analyzed and sorted out. Certain rules and internal relationships among different pathways have been further clarified; this is expected to provide valuable clues for further research. The Wnt/β-catenin pathway is closely associated with the formation of cancer radioresistance, which may be a target for improving the effects of radiotherapy

Advances in Research on the Effects of Wnt Pathway on Macrophage Function in Different Tissues / 中国药学杂志

Wnt protein is a group of highly conserved secretory signal molecules that regulate intercellular signal transduction during embryogenesis by autocrine or paracrine.Wnt signaling pathway is divided into β-catenin-dependent classical signaling pathway and non-classical signaling pathway, which is involved in cell proliferation, migration and differentiation. Macrophages are the main Wnt-secreting cells which are widely distributed in various tissues and participate in the development of many diseases. It is not clear how the Wnt signaling pathway plays a role in the development of macrophages.This article reviews the effects of Wnt signaling pathway on macrophage function in different tissues, and provides a theoretical basis for the diagnosis and treatment of related diseases.

Study regarding the parent-of-origin effect of WNT pathway genes on non-syndromic cleft lip with or without cleft palate among the Chinese population / 中华流行病学杂志

Objective@#Non-syndromic cleft lip with or without cleft palate (NSCL/P) is a common birth defect with its genetic evidence widely explored. This study explored the potential the parent-of-origin (PoO) effect of WNT pathway on the risks of NSCL/P, using a case-parent trio design.@*Methods@#Data on the single nucleotide polymorphism (SNP) of WNT genes were selected from a genome-wide association study (GWAS). A total of 806 Chinese non-syndromic cleft lip patients, with or without cleft palate (NSCL/P) case-parent trios, were gathered from an international consortium. PoO effect of WNT pathway genes and its haplotypes were explored by log-linear models. Additional Wald tests were performed to assess: a) the heterogeneity of PoO effect between different maternal exposures, b) the interaction between PoO effect, c) maternal exposure to environmental tobacco smoke (ETS), and d) multivitamin supplementation during pregnancy. The threshold for statistical significance was adjusted as 3.47×10-4, according to Bonferroni correction.@*Results@#After quality control, a total of 144 SNPs within seven genes were included for analyses, among which 8 SNPs were of potential PoO effect (P<0.05). However, none of them achieved the statistical significance after Bonferroni correction. The haplotype rs4074668-rs12725747 (T-A) on WNT9A showed significant PoO effect, based on the haplotype test for PoO (P=2.74×10-4). In addition, no statistically significant interaction was found in further exploration of this haplotype under environmental exposures as ETS or multivitamin supplementation.@*Conclusions@#Genes in the WNT pathway may influence the NSCL/P risks through the potential PoO effect. Particularly, the haplotype rs4074668-rs12725747 (T-A) on WNT9A presented significant PoO effect on NSCL/P, statistically. From this current study, findings on WNT pathway related risks among the NSCL/P, need to be further validated by independent samples in the future.

Effects of liraglutide on bone metabolism and Wnt pathway in type 2 diabetic rats with osteoporosis / 中华内分泌外科杂志

Objective@#To study the effects of liraglutide on bone metabolism and Wnt pathway in type 2 diabetic osteoporosis rats.@*Methods@#SD rats were randomly divided into control group, model group and liraglutide group. The latter two groups were fed with high-fat and high-sugar diet and intraperitoneally injected with low-dose streptozotocin to establish type 2 diabetic model. Liraglutide group was subcutaneously injected with 0.6 mg/kg/d liraglutide for 8 weeks. Bone mineral density, calcium and phosphorus content, the expression of Wnt pathway molecule [Wnt3a, low density lipoprotein receptor-related protein 5 (LRP5) , β-catenin] and the contents of bone metabolism indicators [ALP, osteocalcin (OC) , osteoprotegerin (OPG) , receptor activator of nuclear factor-κ B ligand (RANKL) , tartrate-resistant acid phosphatase (TrACP) , cross-linked carboxy-terminal telopeptide of type I collagen (CTX-1) ] in serum were determined.@*Results@#The tibial bone mineral density[left (0.158±0.024) vs (0.232±0.041) g/cm2, right (0.152±0.027) vs (0.219±0.038) g/cm2, P<0.05) , the content of calcium and phosphorus [ (5.12±0.58) vs (5.93±0.74) mol/g, (2.93±0.41) vs (3.84±0.56) mol/g, P<0.05) , the expression of Wnt3a, LRP5, β-catenin in tibia [ (0.29±0.06) vs (0.78±0.13) , (0.30±0.05) vs (0.73±0.14) , (0.38±0.06) vs (1.01±0.18) , P<0.05], the content of ALP, OC and OPG in serum of model group were significantly lower than those of control group[ (40.27±7.52) vs (59.29±9.19) ng/ml, (252.45±42.95) vs (341.28±52.39) pg/ml, (0.40±0.06) vs (0.78±0.09) ng/ml, P<0.05) , and the contents of RANKL, TrACP and CTX-1 in serum were significantly higher than those of control group [ (17.79±2.84) vs (12.46±2.09) pg/ml, (56.45±7.79) vs (41.38±8.19) μmol/l, (19.26±3.14) vs (11.39±2.25) pg/ml, P<0.05]; the tibial bone mineral density, the content of calcium and phosphorus, the expression of Wnt3a, LRP5, β-catenin in tibia, the content of ALP, OC and OPG in serum of liraglutide group were significantly higher than those of control group, and the contents of RANKL, TrACP and CTX-1 in serum were significantly lower than those of control group.@*Conclusion@#Liraglutide can improve bone mineral density and bone metabolism in type 2 diabetic rats with osteoporosis, which may be related to activation of Wnt pathway.

Research progress on mechanism and signaling pathway regulation of Chinese materia medica on asthma / 中草药

Asthma is a chronic airway inflammation caused by a variety of factors, involving a variety of cells and cellular components, which is often accompanied by airway hyperresponsiveness and airway remodeling. At present, asthma has become a common chronic respiratory disease. There exists close relationship between the asthma and signaling pathways. MAPK, PI3K/Akt, NF-κB, TGF-β, Notch and Wnt pathways related to the asthma were extensively studied currently. Traditional Chinese medicine shows advantages in the treatment of chronic diseases such as asthma. In order to explain the advantages of traditional Chinese medicine in the treatment of asthma and develop new asthma drugs originated from traditional Chinese medicine, the role of traditional Chinese medicine in the intervention of related signaling pathways of asthma was reviewed. In addition, the relationship between signaling pathway and pathogenesis (airway inflammation, airway hyperresponsive, and airway remodeling) were also discussed.

Effects of liraglutide on bone metabolism and Wnt pathway in type 2 diabetic rats with osteoporosis / 中华内分泌外科杂志

Objective To study the effects of liraglutide on bone metabolism and Wnt pathway in type 2 diabetic osteoporosis rats. Methods SD rats were randomly divided into control group, model group and liraglu-tide group. The latter two groups were fed with high-fat and high-sugar diet and intraperitoneally injected with low-dose streptozotocin to establish type 2 diabetic model. Liraglutide group was subcutaneously injected with 0.6 mg/kg/d liraglutide for 8 weeks. Bone mineral density, calcium and phosphorus content, the expression of Wnt path-way molecule [Wnt3a, low density lipoprotein receptor-related protein 5 (LRP5), β-catenin] and the contents of bone metabolism indicators [ALP, osteocalcin(OC), osteoprotegerin(OPG), receptor activator of nuclear factor-κ B ligand(RANKL), tartrate-resistant acid phosphatase(TrACP), cross-linked carboxy-terminal telopeptide of type I collagen (CTX-1)] in serum were determined. Results The tibial bone mineral density [left (0.158±0.024) vs (0.232±0.041) g/cm2, right(0.152±0.027) vs(0.219±0.038) g/cm2, P<0.05), the content of calcium and phospho-rus [(5.12±0.58) vs (5.93±0.74) mol/g, (2.93±0.41) vs (3.84±0.56) mol/g, P<0.05), the expression of Wnt3a, LRP5, β-catenin in tibia [(0.29±0.06) vs (0.78±0.13), (0.30±0.05) vs (0.73±0.14), (0.38±0.06) vs (1.01± 0.18), P<0.05], the content of ALP, OC and OPG in serum of model group were significantly lower than those of control group [(40.27±7.52) vs (59.29±9.19) ng/ml, (252.45±42.95) vs (341.28±52.39) pg/ml, (0.40±0.06) vs (0.78±0.09) ng/ml, P<0.05), and the contents of RANKL, TrACP and CTX-1 in serum were significantly higher than those of control group [(17.79±2.84) vs(12.46±2.09) pg/ml, (56.45±7.79) vs (41.38±8.19)μmol/l, (19.26± 3.14) vs (11.39±2.25) pg/ml, P<0.05]; the tibial bone mineral density, the content of calcium and phosphorus, the expression of Wnt3a, LRP5, β-catenin in tibia, the content of ALP, OC and OPG in serum of liraglutide group were significantly higher than those of control group, and the contents of RANKL, TrACP and CTX-1 in serum were significantly lower than those of control group. Conclusion Liraglutide can improve bone mineral density and bone metabolism in type 2 diabetic rats with osteoporosis, which may be related to activation of Wnt pathway.

Strawberry Notch 1 (SBNO1) promotes proliferation of spermatogonial stem cells via the noncanonical Wnt pathway in mice / 亚洲男科学杂志(英文版)

While it is known that spermatogonial stem cells (SSCs) initiate the production of male germ cells, the mechanisms of SSC self-renewal, proliferation, and differentiation remain poorly understood. We have previously identified Strawberry Notch 1 (SBNO1), a vertebrate strawberry notch family protein, in the proteome profile for mouse SSC maturation and differentiation, revealing SBNO1 is associated with neonatal testicular development. To explore further the location and function of SBNO1 in the testes, we performed Sbno1 gene knockdown in mice to study the effects of SBNO1 on neonatal testicular and SSC development. Our results revealed that SBNO1 is required for neonatal testicular and SSC development in mice. Particularly, in vitro Sbno1 gene knockdown with morpholino oligonucleotides caused a reduction of SSCs and inactivation of the noncanonical Wnt pathway, through Jun N-terminal kinases. Our study suggests SBNO1 maintains SSCs by promoting the noncanonical Wnt pathway.

Regulation of Wnt signaling pathway on calcium hydroxide-promoted osteogenic differentiation of hDPSCs / 国际生物医学工程杂志

Objective To investigate the role of Wnt/β-catenin signaling pathway in the osteogenic differentiation of human dental pulp stem cells (hDPSCs). Methods After 14 days of the calcium hydroxide training, the cytoskeletal changes of hDPSCs, the expression of β-catenin, i.e. the key promoter of in the Wnt signaling pathway, and the cell localization were detected by laser scanning confocal technique. The Wnt signaling pathways were up-regulated and inhibited, and the osteogenic differentiation and mineralization of hDPSCs were detected by Western Blot and alizarin red staining after 14 days of the training. Results The cytoskeleton of hDPSCs was rearranged by the effect of calcium hydroxide, and theβ-catenin migration from nucleus to cytoplasm were observed. The number of calcium nodules in hDPSCs was decreased after blocking Wnt signaling pathway by Dickkopf-related protein 1 (DKK-1). The calcium hydroxide treatment can promoted dentin sialophosphoprotein (DSPP) expression in hDPSCs. Conclusions Calcium hydroxide can down-regulate the expression of canonical Wnt signaling pathway and promote osteogenic differentiation and mineralization and odontogenetic differentiation of hDPSCs.

Strawberry Notch 1 (SBNO1) promotes proliferation of spermatogonial stem cells via the noncanonical Wnt pathway in mice / 亚洲男科学杂志(英文版)

While it is known that spermatogonial stem cells (SSCs) initiate the production of male germ cells, the mechanisms of SSC self-renewal, proliferation, and differentiation remain poorly understood. We have previously identified Strawberry Notch 1 (SBNO1), a vertebrate strawberry notch family protein, in the proteome profile for mouse SSC maturation and differentiation, revealing SBNO1 is associated with neonatal testicular development. To explore further the location and function of SBNO1 in the testes, we performed Sbno1 gene knockdown in mice to study the effects of SBNO1 on neonatal testicular and SSC development. Our results revealed that SBNO1 is required for neonatal testicular and SSC development in mice. Particularly, in vitro Sbno1 gene knockdown with morpholino oligonucleotides caused a reduction of SSCs and inactivation of the noncanonical Wnt pathway, through Jun N-terminal kinases. Our study suggests SBNO1 maintains SSCs by promoting the noncanonical Wnt pathway.

Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell line

BACKGROUND: Ovarian cancer is a significant cancer-related cause of death in women worldwide. The most used chemotherapeutic regimen is based on carboplatin (CBDCA). However, CBDCA resistance is the main obstacle to a better prognosis. An in vitro drug-resistant cell model would help in the understanding of molecular mechanisms underlying this drug-resistance phenomenon. The aim of this study was to characterize cellular and molecular changes of induced CBDCA-resistant ovarian cancer cell line A2780. METHODS: The cell selection strategy used in this study was a dose-per-pulse method using a concentration of 100 µM for 2 h. Once 20 cycles of exposure to the drug were completed, the cell cultures showed a resistant phenotype. Then, the ovarian cancer cell line A2780 was grown with 100 µM of CBDCA (CBDCA-resistant cells) or without CBDCA (parental cells). After, a drug sensitivity assay, morphological analyses, cell death assays and a RNA-seq analysis were performed in CBDCA-resistant A2780 cells. RESULTS: Microscopy on both parental and CBDCA-resistant A2780 cells showed similar characteristics in morphology and F-actin distribution within cells. In cell-death assays, parental A2780 cells showed a significant increase in phosphatidylserine translocation and caspase-3/7 cleavage compared to CBDCA-resistant A2780 cells (P < 0.05 and P < 0.005, respectively). Cell viability in parental A2780 cells was significantly decreased compared to CBDCA-resistant A2780 cells (P < 0.0005). The RNA-seq analysis showed 156 differentially expressed genes (DEGs) associated mainly to molecular functions. CONCLUSION: CBDCA-resistant A2780 ovarian cancer cells is a reliable model of CBDCA resistance that shows several DEGs involved in molecular functions such as transmembrane activity, protein binding to cell surface receptor and catalytic activity. Also, we found that the Wnt/3-catenin and integrin signaling pathway are the main metabolic pathway dysregulated in CBDCA-resistant A2780 cells.

Canonical Wnt pathway gene expression and their clinical correlation in oral squamous cell carcinoma

Aim: The aim of this study is to explore the prognostic significance and clinicopathological correlations of the Wnt pathway genes in a cohort of surgically treated patients with oral squamous cell carcinoma (OSCC) patients. Settings and Design: A prospective genetic study on patients with OSCC was carried out during the period from July 2014 to January 2016. Informed consent from patients and institutional ethical approval for the study was obtained and the guidelines were strictly followed for collection of samples. Subjects and Methods: Clinical data and mRNA expression analysis of ten genes in the canonical Wnt pathway were evaluated and their relationships with clinical and demographic variables were studied in 58 tissue samples. Wnt-3a, β-catenin, secreted frizzled-related proteins sFRP-1, sFRP-2, sFRP-4, sFRP-5, Wnt inhibitory factor 1, dickkopf-1, c-MYC, and cyclin-D1 from cancer (n = 29) and normal (n = 29) tissue samples were investigated using quantitative reverse transcription-polymerase chain reaction. Statistical Analysis: Descriptive statistics were used to summarize the sample characteristics and clinical variables. If the data were normal, then parametric tests were used; otherwise, nonparametric alternatives were used. All the analyses were carried out using SPSS version 23.0 (IBM SPSS Inc., USA). Results: Expression of sFRP-1, sFRP-2, and sFRP-5 in control samples and expression of c-MYC and cyclin D1 in cancer samples showed statistical significance. Significant expression of Wnt3A was observed among patients who had recurrence and were deceased. Conclusion: Wnt3A, β-catenin, and cyclin D1 are recognized as key components of Wnt/β-catenin signaling. However, in this study, there was no significant expression of all the three genes in OSCC. The proto-oncogene c-MYC showed statistically significant upregulation in cancer tissue samples suggesting that the OSCC among South Indian population is primarily not mediated by the canonical Wnt signaling pathway.

Strontium ranelate inhibits wear particle-induced aseptic loosening in mice

The imbalance between bone formation and osteolysis plays a key role in the pathogenesis of aseptic loosening. Strontium ranelate (SR) can promote bone formation and inhibit osteolysis. The aim of this study was to explore the role and mechanism of SR in aseptic loosening induced by wear particles. Twenty wild-type (WT) female C57BL/6j mice and 20 sclerostin-/- female C57BL/6j mice were used in this study. Mice were randomly divided into four groups: WT control group, WT SR group, knockout (KO) control group, and KO SR group. We found that SR enhanced the secretion of osteocalcin (0.72±0.007 in WT control group, 0.98±0.010 in WT SR group, P=0.000), Runx2 (0.34±0.005 in WT control group, 0.47±0.010 in WT SR group, P=0.000), β-catenin (1.04±0.05 in WT control group, 1.22±0.02 in WT SR group, P=0.000), and osteoprotegerin (OPG) (0.59±0.03 in WT control group, 0.90±0.02 in WT SR group, P=0.000). SR significantly decreased the level of receptor activator for nuclear factor-κB ligand (RANKL) (1.78±0.08 in WT control group, 1.37±0.06 in WT SR group, P=0.000) and improved the protein ratio of OPG/RANKL, but these effects were not observed in sclerostin-/- mice. Our findings demonstrated that SR enhanced bone formation and inhibited bone resorption in a wear particle-mediated osteolysis model in wild-type mice, and this effect relied mainly on the down-regulation of sclerostin levels to ameliorate the inhibition of the canonical Wnt pathway.

Key signal molecules of wingless-related integration pathway and targeted therapy for liver cancer / 中华肝脏病杂志

Hepatocellular carcinoma is one of the most common digestive system tumors. Its occurrence and development are considered as multi-factorial and multi-step process. Recent studies have shown that wingless-related integration (Wnt) pathway plays an important role in the HCC progression and is associated with malignant transformation of hepatocytes, HCC metastasis, drug resistance and liver cancer stem cells. This article analyzes the expression of key signaling molecules in Wnt pathway and its value in diagnosis, prognosis and targeted therapy, and outlines the research progress of Wnt pathway targeted drugs for the treatment of HCC, with a view to providing targeted therapy research for HCC reference.

CpG Island Methylator Phenotype and Methylation of Wnt Pathway Genes Together Predict Survival in Patients with Colorectal Cancer

PURPOSE: Dysregulation of the Wnt pathway is a crucial step in the tumorigenesis of colorectal cancer (CRC). This study aimed to determine whether DNA methylation of Wnt pathway genes helps predict treatment response and survival in patients with metastatic or recurrent CRC. MATERIALS AND METHODS: We retrospectively collected primary tumor tissues from 194 patients with metastatic or recurrent CRC. Pyrosequencing was used to examine the methylation of 10 CpG island loci in DNA extracted from formalin-fixed paraffin-embedded specimens. To elucidate the predictive role of DNA methylation markers, Kaplan-Meier survival estimation and Cox regression were performed for progression-free survival and overall survival (OS). RESULTS: The methylation frequencies of the 10 genes analyzed (p16, p14, MINT1, MINT2, MINT31, hMLH1, DKK3, WNT5A, AXIN2, and TFAP2E) were 47.9%, 10.8%, 21.1%, 16.0%, 20.6%, 0.5%, 53.1%, 32.0%, 2.6%, and 2.1%, respectively. We divided patients into three groups based on the number of methylated genes (group 1, no methylation n=38; group 2, 1–2 methylations n=92; group 3, 3 or more methylations n=64). Among patients treated with palliative chemotherapy (n=167), median OSs of groups 1, 2, and 3 were 39.1, 39.7, and 29.1 months, respectively (log rank p=0.013). After adjustment, number of methylations was identified as an independent poor prognostic factor (0–2 methylated vs. ≥3 methylated: hazard ratio, 1.72; 95% confidence interval, 1.16–2.56, p=0.007). CONCLUSION: This study suggests that methylation of Wnt pathway genes, in addition to known CpG island methylator phenotype markers, may help predict treatment outcome and survival in patients with CRC.

K (lysine) acetyltransferase 2A affects the osteogenic differentiation of periodontal ligament stem cells through the canonical Wnt pathway / 华西口腔医学杂志

OBJECTIVE@#This study aims to investigate the mechanism of K (lysine) acetyltransferase 2A (KAT2A) regulation and control on the osteogenic differentiation of periodontal ligament stem cells (PDLSCs).@*METHODS@#The expression levels of KAT2A in PDLSCs were compared from each generation of the normal (H-PDLSCs) and periodontitis tissues (P-PDLSCs). The influences of KAT2A gene interference on the osteogenic differentiation of PDLSCs were also detected. In addition, the influences of the KAT2A gene interference to the canonical Wnt pathway and ligands were detected. The upstream and down-stream relationships between KAT2A and canonical Wnt pathway were also determined.@*RESULTS@#The decreased expression of KAT2A in PDLSCs from the inflammatory tissue in each generation was compared with that in PDLSCs from the healthy tissue, and the difference was statistically significant (P<0.05). When the KAT2A gene was disrupted, the osteogenesis ability of PDLSC was declined, and the difference was statistically significant (P<0.05). The canonical Wnt pathway was activated, and the antagonist Dickkopf-1 (DKK-1) was reduced. After the DKK-1 addition, the osteogenic differentiation of the disturbed PDLSCs was recovered, and KAT2A was unaffected.@*CONCLUSIONS@#The KAT2A expression in PDLSCs was decreased because of perio-dontitis. The classical Wnt pathway was activated to inhibit the osteogenic differentiation of the cells.

Effects of Wnt pathway inhibitor ETC-159 on proliferation and migration of OSCC cells / 实用肿瘤学杂志

Objective The objective of this study was to investigate the effect of Wnt pathway inhibitor-ETC-159 on pro-liferation and migration of oral squamous cell carcinoma(SCC-15)cells and to explore its mechanism.Methods SCC-15 cells were treated with DMSO and ETC-159 for 12 h or 24 h.Cell proliferation was detected by CCK8 kit.Transwell assay was used to de-tect the ability of cell migration.Western blotting was used to detect cell migration related to proteins i.e.Wnt3a and β-catenin,pro-liferation and migration related proteins i.e.c-Myc,cyclin D1,CD146.Results After treated with ETC-159 for 12 or 24 h,the proliferation,migration and expression of Wnt3a,β-catenin,c-Myc,cyclin D1 and CD146 in SCC-15 cells were significantly de-creased when compared to the DMSO group(P<0.05).Conclusion Wnt signaling pathway inhibitor-ETC-159 can inhibit the proliferation and migration of SCC-15 cells by decreased levels of c-Myc,cyclin D1 and CD146.