Hepatocellular carcinoma(HCC),a common malignancy worldwide,still lacks effective clinical treatment.The study aimed to investigate the oncogenes that affect the progression of HCC and their possible mechanisms.In our...Hepatocellular carcinoma(HCC),a common malignancy worldwide,still lacks effective clinical treatment.The study aimed to investigate the oncogenes that affect the progression of HCC and their possible mechanisms.In our study,we initially confirmed a higher level of PRDX2 in the bile of HCC patients compared to those with choledocholithiasis by 2-DE,LC-MS,and ELISA.Subsequently,we demonstrated the high expression of peroxiredoxin 2(PRDX2)in HCC based on the TCGA database and clinical sample analysis.Furthermore,PRDX2 overexpression enhanced the viability of HCC cells.And PRDX2 silencing induced senescence of HCC cells.In vivo,knockdown of PRDX2 significantly reduced the weight of xenograft tumors.PRDX2 also was found to activate the Wnt/β-catenin pathway by inducingβ-catenin nuclear translocation.Consequently,we proved that silencing PRDX2 could inhibit proliferation and Wnt/β-catenin pathway while promoting senescence in HCC cells.展开更多
An important factor in the emergence and progre sion of osteosarcoma(OS)is the dysregulated expression of microRNAs(miRNAs).Transcription factor 7-like 1(TCF7LI),a member of the T cell factor/lymphoid enhancer factor(...An important factor in the emergence and progre sion of osteosarcoma(OS)is the dysregulated expression of microRNAs(miRNAs).Transcription factor 7-like 1(TCF7LI),a member of the T cell factor/lymphoid enhancer factor(TCF/LEF)transcription factor family,interacts with the Wnt signaling pathway regulator β-catenin and acts as a DNA-specific binding protein.This study sought to elucidate the impact of the interaction between miR 3293p and TCF7L1 on.the growth and apoptosis of OS and analyze the regulatory expression relationship between miRNA and mRNA in osteosarcoma cells using a variety of approaches.MiR329-3p was significantly downregulated,while TCF7L1 was considerably up-regulated in all examined OS cell lines.Additionally,a clinical comparison study was performed using the TCGA database.Subsequently,the regulatory relationship between miR-329-3p and TCF7L1 on the proliferation and apoptosis of OS cells was verified through in vitro and in vivo experiments.When miR 329-3p was transfected into the OS cell line,the expression of TCF7L1 decreased,the proliferation of OS cells was inhibited,the cytoskeleton disintegrated,and the nucleus condensed to fom apoptotic bodies.The expression of proteins that indicate apoptosis increased simultaneously.The cell cycle was arrested in the G0/G1 phase,and the G1/S transition was blocked.The introduction of miR 3293p also inhibited downstream Cyclin D1 of the Wnt pathway.Xenograf experiments indicated that the overexpression of miR-329-3p signi ficanly inhibited the growth of OS xenografts in nude mice,and the expression of TCF7L1 and C-Myc in tumor tssues decreased.MiR 329-3p was significantly reduced in OS cells and played a suppressive role in tumorigenesis and proliferation by targeting TCF7L1 both in vitro and in vivo.Osteosarcoma cell cycle arrest and pathway inhibition were observed upon the regulation of TCF7LI by miR 3293p.Summarizing these results,it can be inferred that miR.3293p exerts anticancer efects in osteosarcoma by inhibiting TCF7L1.展开更多
Increasing the osteogenic differentiation ability and decreasing the adipogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs)is a potential strategy for the treatment of osteoporosis(OP).Natural...Increasing the osteogenic differentiation ability and decreasing the adipogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs)is a potential strategy for the treatment of osteoporosis(OP).Naturally derived oligosaccharides have shown significant anti-osteoporotic effects.Nystose(NST),an oligosaccharide,was isolated from the roots of Morinda officinalis How.(MO).The aim of the present study was to investigate the effects of NST on bone loss in ovariectomized mice,and explore the underlying mechanism of NST in promoting differentiation of BMSCs to osteoblasts.Administration of NST(40,80 and 160 mg/kg)and the positive control of estradiol valerate(0.2 mg/kg)for 8 weeks significantly prevented bone loss induced by ovariectomy(OVX),increased the bone mass density(BMD),improved the bone microarchitecture and reduced urine calcium and deoxypyridinoline(DPD)in ovariectomized mice,while inhibited the increase of body weight without significantly affecting the uterus weight.Furthermore,we found that NST increased osteogenic differentiation,inhibited adipogenic differentiation of BMSCs in vitro,and upregulated the expression of the key proteins of BMP and Wnt/β-catenin pathways.In addition,Noggin and Dickkopf-related protein-1(DKK-1)reversed the effect of NST on osteogenic differentiation and expression of the key proteins in BMP and Wnt/β-catenin pathway.The luciferase activities and the molecular docking analysis further supported the mechanism of NST.In conclusion,these results indicating that NST can be clinically used as a potential alternative medicine for the prevention and treatment of postmenopausal osteoporosis.展开更多
Background:Actinidia chinensis Planch.roots(AcRoots)have been applied as an anti-inflammatory and antitumor drug in the treatment of gastric cancer(GC).However,their mechanisms against GC cells remain unclear.To inves...Background:Actinidia chinensis Planch.roots(AcRoots)have been applied as an anti-inflammatory and antitumor drug in the treatment of gastric cancer(GC).However,their mechanisms against GC cells remain unclear.To investigate the anticancer effect of AcRoots in GC and the possible underlying mechanism by using network pharmacology.Methods:Differentially expressed genes between gastric precancerous lesions and cancer were analyzed in Gene Expression Omnibus datasets,and these genes were overlapped with potential targets of AcRoots.Potential targets and pathways for AcRoots treatment of GC predicted by network pharmacology.Furthermore,we used the GC cell line HGC27 to explore the molecular mechanisms in the context of hub genes in apoptosis,invasion,metastasis,and epithelial to mesenchymal transition-promoting factors.Molecular docking between hub targets and active drug components was also performed.Results:Network pharmacological analysis suggested that the potential mechanism was related to the Wnt pathway and predicted nine hub genes.In in vitro studies,AcRoots significantly decreased HGC27 cell viability and promoted apoptosis by upregulating caspase3 and downregulating Bcl2.Moreover,it suppressed invasion and metastasis as well as the expression of epithelial to mesenchymal transition-related factors.In addition,AcRoots affected the phosphorylation level of GSK3β(Ser9)in the Wnt pathway to promote the degradation ofβ-catenin,resulting in the downregulation of the downstream target genes c-myc,cyclin D1 and snail.All the experimental results were consistent with the network pharmacology results.Conclusion:This study combined network pharmacology with in vitro experiments to provide valid evidence for the clinical promotion of AcRoots.展开更多
BACKGROUND Exosomal miRNAs play crucial roles in many central nervous system diseases.Cerebral small vessel disease(CVSD)is a small vessel disease that is affected by various factors.This study aimed to investigate th...BACKGROUND Exosomal miRNAs play crucial roles in many central nervous system diseases.Cerebral small vessel disease(CVSD)is a small vessel disease that is affected by various factors.This study aimed to investigate the role of exosomal miR-320e in the Wnt/β-catenin pathway stimulated by oxidative stress and assess its clinical correlation with psychiatric symptoms in patients with CVSD.AIM To explore whether exosomal miR-320e could suppress the Wnt/β-catenin pathway and play a protective role in CVSD progression,as well as examine its potential correlation with cognitive impairment and depression in patients with CVSD.METHODS Differentially expressed exosomal miRNAs were filtered by sequencing plasma exosomes from patients with CVSD and healthy controls.Bioinformatics and dual luciferase analyses were used to confirm the binding of miR-320e to Wnt2,and the mRNA and protein levels of downstream components in the Wnt/β-catenin pathway were evaluated when overexpressed or with knockdown of miR-320e under H2O2-induced oxidative stress.In addition,Wnt2-targeting siRNA was used to confirm the role of miR-320e in the Wnt2-mediated inhibition of the Wnt/β-catenin pathway.A retrospective analysis was conducted among patients with CVSD to confirm the correlation between miR-320e expression and the severity of cognitive impairment and depression,which were quantified using the Montreal Cognitive Assessment(MoCA)/Executive Function Assessment(EFA),and the Hamilton Depression Scale(HAMD)/Beck Depression Inventory(BDI),respectively.RESULTS High-throughput sequencing revealed that exosomal miR-320e was downregulated in patients with CVSD.Bioinformatics analysis and dual-luciferase reporter gene experiments showed that exosomal miR-320e inhibited the Wnt/β-catenin pathway in response to oxidative stress by targeting the 3'noncoding region of Wnt2.Uptake of exosomes carrying miR-320e into endothelial cells could also target Wnt2 and inhibit the Wnt2/β-catenin pathway.Elevated miR-320e expression may protect patients with CVSD from relatively severe cognitive impairment and depression,as it was found to have a positive correlation with the MoCA/EFA and HAMD/BDI scores.CONCLUSION Our results suggest that exosomal miR-320e suppresses the Wnt/β-catenin pathway and may play a protective role in CVSD progression.展开更多
BACKGROUND Hepatic fibrosis is a common pathological process of chronic liver diseases with various causes,which can progress to cirrhosis.AIM To evaluate the effect and mechanism of action annexin(Anx)A1 in liver fib...BACKGROUND Hepatic fibrosis is a common pathological process of chronic liver diseases with various causes,which can progress to cirrhosis.AIM To evaluate the effect and mechanism of action annexin(Anx)A1 in liver fibrosis and how this could be targeted therapeutically.METHODS CCl4(20%)and active N-terminal peptide of AnxA1(Ac2-26)and N-formylpeptide receptor antagonist N-Boc-Phe-Leu-Phe-Leu-Phe(Boc2)were injected intraperitoneally to induce liver fibrosis in eight wild-type mice/Anxa1 knockout mice,and to detect expression of inflammatory factors,collagen deposition,and the role of the Wnt/β-catenin pathway in hepatic fibrosis.RESULTS Compared with the control group,AnxA1,transforming growth factor(TGF)-β1,interleukin(IL)-1βand IL-6 expression in the liver of mice with hepatic fibrosis induced by CCl4 was significantly increased,which promoted collagen deposition and expression ofα-smooth muscle actin(α-SMA),collagen type I and connective tissue growth factor(CTGF),and increased progressively with time.CCl4 induced an increase in TGF-β1,IL-1βand IL-6 in liver tissue of AnxA1 knockout mice,and the degree of liver inflammation and fibrosis and expression ofα-SMA,collagen I and CTGF were significantly increased compared with in wild-type mice.After treatment with Ac2-26,expression of liver inflammatory factors,degree of collagen deposition and expression of a-SMA,collagen I and CTGF were decreased compared with before treatment.Boc2 inhibited the anti-inflammatory and antifibrotic effects of Ac2-26.AnxA1 downregulated expression of the Wnt/β-catenin pathway in CCl4-induced hepatic fibrosis.In vitro,lipopolysaccharide(LPS)induced hepatocyte and hepatic stellate cell(HSC)expression of AnxA1.Ac2-26 inhibited LPS-induced RAW264.7 cell activation and HSC proliferation,decreased expression ofα-SMA,collagen I and CTGF in HSCs,and inhibited expression of the Wnt/β-catenin pathway after HSC activation.These therapeutic effects were inhibited by Boc2.CONCLUSION AnxA1 inhibited liver fibrosis in mice,and its mechanism may be related to inhibition of HSC Wnt/β-catenin pathway activation by targeting formylpeptide receptors to regulate macrophage function.展开更多
BACKGROUND Mesenchymal stem cells(MSCs)have been applied to treat degenerative articular diseases,and stromal cell-derived factor-1α(SDF-1α)may enhance their therapeutic efficacy.However,the regulatory effects of SD...BACKGROUND Mesenchymal stem cells(MSCs)have been applied to treat degenerative articular diseases,and stromal cell-derived factor-1α(SDF-1α)may enhance their therapeutic efficacy.However,the regulatory effects of SDF-1αon cartilage differentiation remain largely unknown.Identifying the specific regulatory effects of SDF-1αon MSCs will provide a useful target for the treatment of degenerative articular diseases.AIM To explore the role and mechanism of SDF-1αin cartilage differentiation of MSCs and primary chondrocytes.METHODS The expression level of C-X-C chemokine receptor 4(CXCR4)in MSCs was assessed by immunofluorescence.MSCs treated with SDF-1αwere stained for alkaline phosphatase(ALP)and with Alcian blue to observe differentiation.Western blot analysis was used to examine the expression of SRY-box transcription factor 9,aggrecan,collagen II,runt-related transcription factor 2,collagen X,and matrix metalloproteinase(MMP)13 in untreated MSCs,of aggrecan,collagen II,collagen X,and MMP13 in SDF-1α-treated primary chondrocytes,of glycogen synthase kinase 3β(GSK3β)p-GSK3βandβ-catenin expression in SDF-1α-treated MSCs,and of aggrecan,collagen X,and MMP13 in SDF-1α-treated MSCs in the presence or absence of ICG-001(SDF-1αinhibitor).RESULTS Immunofluorescence showed CXCR4 expression in the membranes of MSCs.ALP stain was intensified in MSCs treated with SDF-1αfor 14 d.The SDF-1αtreatment promoted expression of collagen X and MMP13 during cartilage differentiation,whereas it had no effect on the expression of collagen II or aggrecan nor on the formation of cartilage matrix in MSCs.Further,those SDF-1α-mediated effects on MSCs were validated in primary chondrocytes.SDF-1αpromoted the expression of p-GSK3βandβ-catenin in MSCs.And,finally,inhibition of this pathway by ICG-001(5μmol/L)neutralized the SDF-1α-mediated up-regulation of collagen X and MMP13 expression in MSCs.CONCLUSION SDF-1αmay promote hypertrophic cartilage differentiation in MSCs by activating the Wnt/β-catenin pathway.These findings provide further evidence for the use of MSCs and SDF-1αin the treatment of cartilage degeneration and osteoarthritis.展开更多
The activation of the Wnt/β-catenin signaling pathway in oval cells after liver injury is implicated in hepatocarcinogenesis.Diwu Yanggan capsule is a Chinese herbal medicine that has been used for treating liver dis...The activation of the Wnt/β-catenin signaling pathway in oval cells after liver injury is implicated in hepatocarcinogenesis.Diwu Yanggan capsule is a Chinese herbal medicine that has been used for treating liver disorder.The present study aimed to examine the mechanism by which Diwu Yanggan inhibits liver carcinogenesis,and the involvement of the Wnt/β-catenin signaling pathway.Diwu Yanggan capsule was administered to 2-acetaminofluorene/partial hepatectomy(2-AAF/PH)rats,a murine model of liver injury.The biomarkers of oval cells and key proteins in the Wnt/p-catenin signaling pathway were assessed on postoperative day 8,10,14,17,19 and 22.The results showed that treatment with Diwu Yanggan was associated with reduced expression of oval cell and stem cell biomarkers in the 2-AAF/PH animals.The expression pattern of key proteins in the Wnt/β-catenin pathway was altered in Diwu Yanggan-treated animals,indicating that the Diwu Yanggan treatment accelerated the activation of the Wnt/β-catenin pathway in the initial stage and contributed to its deactivation in the later stage.Histological findings indicated that hepatocyte proliferation was suppressed in Diwu Yanggan-treated animals,compared with untreated 2-AAF/PH animals.Taken together,Diwu Yanggan capsule may reduce the risk of hepatocarcinogenesis by modulating the Wnt/β-catenin signaling pathway.展开更多
The neuropsychiatric disease named obsessive-compulsive disorder is composed by obsessions and/or compulsions.Obsessive-compulsive disorder etiologies are undefined.However,numerous mechanisms in several localizations...The neuropsychiatric disease named obsessive-compulsive disorder is composed by obsessions and/or compulsions.Obsessive-compulsive disorder etiologies are undefined.However,numerous mechanisms in several localizations are implicated.Some studies showed that both glutamate,inflammatory factors and oxidative stress could have main functions in obsessive-compulsive disorder.Glycogen synthase kinase-3β,the major negative controller of the WNT/β-catenin pathway is upregulated in obsessive-compulsive disorder.In obsessive-compulsive disorder,some studies presented the actions of the different circadian clock genes.WNT/β-catenin pathway and circadian clock genes appear to be intricate.Thus,this review focuses on the interaction between circadian clock genes and the WNT/β-catenin pathway in obsessive-compulsive disorder.展开更多
In this study,fucoidans were extracted from the sea cucumber Thelenota ananas(Ta-FUCs)by enzymatic degradation.Four products with molecular weights of 1380.3,524.0,182.4,and 110.3 kDa were obtained,and the Ta-FUC show...In this study,fucoidans were extracted from the sea cucumber Thelenota ananas(Ta-FUCs)by enzymatic degradation.Four products with molecular weights of 1380.3,524.0,182.4,and 110.3 kDa were obtained,and the Ta-FUC showing optimal anti-adipogenic activities was determined.Results of MTT and Oil red O staining analyses showed that the Ta-FUCs inhibited the proliferation and differentiation of 3T3-L1 adipocytes.Futhermore,Ta-FUCs significantly downregulated the key transcriptional factors,such as SREBP-1c,PPARγ,and C/EBPαof adipocytes.The Ta-FUCs also activated Wnt/β-catenin pathway-related genes,such asβ-catenin,LRP5,and FrZ.The Ta-FUCs suppressed lipid accumulation in 3T3-L1 adipocytes possibly by decreasing the expression of genes ACC,FAS,ME,GPAT,DGAT,and PILN,which are important in the synthesis of fatty acids and triglycerides;and by increasing the expression of genes PPARα,CPT-1α,and ACOX,which are crucial in fatty acidβ-oxidation.The anti-adipogenic activities initially increased and then declined with decreasing molecular weight.Among the Ta-FUCs,the 182.4 kDa Ta-FUC exhibited optimal bioactivities.This study reports for the first time that Ta-FUCs can prevent obesity by regulating the differentiation and lipid accumulation of adipocytes.展开更多
Osteoporosis is a frequently occurring bone remodeling disorder worldwide with one characteristic being decreasing bone mineral density and a predisposition to bone fracture,which diminishes patients’quality of life....Osteoporosis is a frequently occurring bone remodeling disorder worldwide with one characteristic being decreasing bone mineral density and a predisposition to bone fracture,which diminishes patients’quality of life.Several studies showed that imbalance between the osteogenesis and adipogenesis of bone marrow mesenchymal stem cells(BMSCs)took part in the development of osteoporosis.In previous study,we found MIR22HG regulated the osteogenesis of human BMSCs positively.In this study,we found that MIR22HG was decreased during the adipogenesis of human BMSCs and exerted negative effects on adipogenesis with the involvement of Wnt/β-catenin signaling pathway both in vitro and in vivo.Nitazoxanide could inhibit Wnt signaling and relieve MIR22HG’s suppression on adipogenesis.These findings indicated that MIR22HG had great potential in clinical application for osteoporosis treatment and prevention.展开更多
Recent studies showed that activation of Wnt/β-catenin pathway promoted the differentiation of osteoblast-like cells in the arterial calcification, but its mechanism remains unknown. In this study, the hypothesis tha...Recent studies showed that activation of Wnt/β-catenin pathway promoted the differentiation of osteoblast-like cells in the arterial calcification, but its mechanism remains unknown. In this study, the hypothesis that Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by upregulating the expression of receptor activator of NF-κB ligand (RANKL) was examined. LiCl was used to activate the Wnt/β-catenin pathway. The differentiation of osteoblast-like cells was observed by Von Kossa staining, calcium content assay, alkaline phosphatase (ALP) activity assay, and detection of osteocalcin expression. Real-time PCR was performed to detect the expression of RANKL and osteoprotegerin (OPG, the decoy receptor of RANKL) during the osteoblast-like cell differentiation. Different concentrations of OPG were added to the culture media respectively to inhibit the function of RANKL, and the change in the differentiation of osteoblast-like cells was evaluated. The results showed that when the Wnt/β-catenin pathway was activated by LiCl, the expression of RANKL was significantly in-creased, which coincided with the differentiation of osteoblast-like cells (P<0.05), and the OPG treatment could partly attenuate the promoting effect of Wnt/β-catenin pathway on the differentiation of osteoblast-like cells (P<0.05), but it failed to completely abolish such effect. It was concluded that activation of Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by both RANKL-dependent and RANKL-independent mechanisms.展开更多
In this study,the hypothesis that Wnt/β-catenin pathway is involved in the arterial calcification by regulating the osteoprotegerin(OPG)/receptor activator of NF-κB ligand(RANKL)system was tested.Theβ-catenin expre...In this study,the hypothesis that Wnt/β-catenin pathway is involved in the arterial calcification by regulating the osteoprotegerin(OPG)/receptor activator of NF-κB ligand(RANKL)system was tested.Theβ-catenin expression was measured in the warfarin-induced calcified arteries and the osteoblast-like cells differentiating from smooth muscle cells(SMCs)by immunohistochemistry and Western blotting.The Wnt/β-catenin pathway was activated or inhibited by lithium chloride(LiCl)or dickkopf 1(DKK1)in vitro and in vivo.Then the calcification level was determined by von Kossa staining,Ca^2+content assay,and alkaline phosphatase(ALP)activity assay.The expression levels of osteocalcin,OPG and RANKL were detected by Western blotting or real-time PCR.The results showed that in calcified arteries and OBL cells,the activation of Wnt/β-catenin pathway significantly enhanced the calcification as evidenced by increased von Kossa stains,Ca^2+contcnts,ALP activities,and osteocalcin expression levels(P<0.05),and it promoted the RANKL expression(P<0.05),but slightly affected the OPG expression.These results indicated that the activation of Wnt/β-catenin pathway worsens the arterial calcification,probably by promoting the RANKL expression.展开更多
Gastric cancer remains one of the most common cancers worldwide and one of the leading cause for cancerrelated deaths. Gastric adenocarcinoma is a multifactorial disease that is genetically, cytologically and architec...Gastric cancer remains one of the most common cancers worldwide and one of the leading cause for cancerrelated deaths. Gastric adenocarcinoma is a multifactorial disease that is genetically, cytologically and architecturally more heterogeneous than other gastrointestinal carcinomas.The aberrant activation of the Wnt/β-catenin signaling pathway is involved in the development and progression of a significant proportion of gastric cancer cases. This review focuses on the participation of the Wnt/b-catenin pathway in gastric cancer by offering an analysis of the relevant literature published in this field. Indeed, it is discussed the role of key factors in Wnt/β-catenin signaling and their downstream effectors regulating processes involved in tumor initiation, tumor growth, metastasis and resistance to therapy. Available data indicate that constitutive Wnt signalling resulting from Helicobacter pylori infection and inactivation of Wnt inhibitors(mainly by inactivating mutations and promoter hypermethylation) play an important role in gastric cancer. Moreover, a number of recent studies confirmed CTNNB1 and APC as driver genes in gastric cancer. The identification of specific membrane, intracellular, and extracellular components of the Wnt pathway has revealed potential targets for gastric cancer therapy. High-throughput "omics" approaches will help in the search for Wnt pathway antagonist in the near future.展开更多
BACKGROUND Our previous study demonstrated that RBBP4 was upregulated in colon cancer and correlated with poor prognosis of colon cancer and hepatic metastasis.However,the potential biological function of RBBP4 in col...BACKGROUND Our previous study demonstrated that RBBP4 was upregulated in colon cancer and correlated with poor prognosis of colon cancer and hepatic metastasis.However,the potential biological function of RBBP4 in colon cancer is still unknown.AIM To investigate the biological role and the potential mechanisms of RBBP4 in colon cancer progression.METHODS Real-time polymerase chain reaction and western blot analysis were used to detect the expression of RBBP4 in colon cancer cell lines.The cell proliferation and viability of SW620 and HCT116 cells with RBBP4 knockdown was detected by Cell Counting Kit-8 and 5-ethynyl-2’-deoxyuridine staining.The transwell assay was used to detect the invasion and migration capabilities of colon cancer cells with RBBP4 knockdown.Flow cytometry apoptosis assay was used to detect the apoptosis of colon cancer cells.Western blotting analysis was used to detect the expression of epithelial-mesenchymal transition and apoptosis related markers in colon cancer.The nuclear translocation ofβ-catenin was examined by Western blotting analysis in colon cancer cells with RBBP4 knockdown.The TOPFlash luciferase assay was used to detect the effect of RBBP4 on Wnt/β-catenin activation.The rescue experiments were performed in colon cancer cells treated with Wnt/β-catenin activator LiCl and RBBP4 knockdown.RESULTS We found that RBBP4 was highly expressed in colon cancer cell lines.The 5-ethynyl-2’-deoxyuridine assay showed that knockdown of RBBP4 significantly inhibited cell proliferation.RBBP4 inhibition reduced cell invasion and migration via regulating proteins related to epithelial-mesenchymal transition.Knockdown of RBBP4 significantly inhibited survivin-mediated apoptosis.Mechanistically,the TOPFlash assay showed that RBBP4 knockdown increased activity of the Wnt/β-catenin pathway.Meanwhile,RBBP4 knockdown suppressed nuclear translocation ofβ-catenin.With Wnt/β-catenin activator,rescue experiments suggested that the role of RBBP4 in colon cancer progression was dependent on Wnt/β-catenin pathway.CONCLUSION RBBP4 promotes colon cancer development via increasing activity of the Wnt/β-catenin pathway.RBBP4 may serve as a novel therapeutic target in colon cancer.展开更多
Colorectal cancer(CRC)continues to be one of the main causes of death from cancer because patients progress unfavorably due to resistance to current therapies.Dysregulation of the Wnt/β-catenin pathway plays a fundam...Colorectal cancer(CRC)continues to be one of the main causes of death from cancer because patients progress unfavorably due to resistance to current therapies.Dysregulation of the Wnt/β-catenin pathway plays a fundamental role in the genesis and progression of several types of cancer,including CRC.In many subtypes of CRC,hyperactivation of theβ-catenin pathway is associated with mutations of the adenomatous polyposis coli gene.However,it can also be associated with other causes.In recent years,studies of the tumor microenvironment(TME)have demonstrated its importance in the development and progression of CRC.In this tumor nest,several cell types,structures,and biomolecules interact with neoplastic cells to pave the way for the spread of the disease.Cross-communications between tumor cells and the TME are then established primarily through paracrine factors,which trigger the activation of numerous signaling pathways.Crucial advances in the field of oncology have been made in the last decade.This Minireview aims to actualize what is known about the central role of the Wnt/β-catenin pathway in CRC chemoresistance and aggressiveness,focusing on crosscommunication between CRC cells and the TME.Through this analysis,our main objective was to increase the understanding of this complex disease considering a more global context.Since many treatments for advanced CRC fail due to mechanisms involving chemoresistance,the data here exposed and analyzed are of great interest for the development of novel and effective therapies.展开更多
Objective: To investigate the effect of arthroscopic debridement combined with intra-articular sodium hyaluronate injection on the Wnt/β-catenin pathway in joint fluid of patients with knee osteoarthritis. Methods: P...Objective: To investigate the effect of arthroscopic debridement combined with intra-articular sodium hyaluronate injection on the Wnt/β-catenin pathway in joint fluid of patients with knee osteoarthritis. Methods: Patients undergoing arthroscopic debridement for knee osteoarthritis in the hospital between June 2014 and April 2017 were selected as the research subjects and randomly divided into the observation group who accepted arthroscopic debridement combined with intra-articular sodium hyaluronate injection and the control group who accepted arthroscopic debridement alone. The Wnt/β-catenin pathway molecules as well as downstream collagen metabolism molecules and bone metabolism molecules in the joint fluid were measured before surgery and 7 d after surgery. Results: 7 d after surgery, Wnt1, Wnt5a, Wnt7b, β-catenin, MMP1, MMP13, CTX-I, CTX-II, ADAMTS4, ADAMTS5, RANK, RANKL and TRACP5b mRNA expression in joint fluid of both groups of patients were higher than those before surgery whereas OPG and OC contents were lower than those before surgery, and Wnt1, Wnt5a, Wnt7b, β-catenin, MMP1, MMP13, CTX-I, CTX-II, ADAMTS4, ADAMTS5, RANK, RANKL and TRACP5b mRNA expression in joint fluid of observation group were significantly lower than those of control group whereas OPG and OC contents were significantly higher than those of control group. Conclusion: Arthroscopic debridement combined with intra-articular sodium hyaluronate injection can inhibit the collagen metabolism and bone metabolism disorder mediated by Wnt/β-catenin after surgery.展开更多
Objective: To observe MET-associated alteration during the trans-differentiation from MSCs to neuron-like cells, and to explore the possible molecular mechanism. Methods: Bone marrow MSCs were isolated from rat femur ...Objective: To observe MET-associated alteration during the trans-differentiation from MSCs to neuron-like cells, and to explore the possible molecular mechanism. Methods: Bone marrow MSCs were isolated from rat femur and purified in continuous cell culture. After induced differentiation to neuron-like cells by the combination of butylated hydroxyanisole(BHA)and dimethyl sulfoxide(DMSO), cells were tested by comparative polymerase chain reaction(PCR) for the relative expression of MET biomarkers and transcription factors, and for cell cycle by flow cytometry. Meanwhile, target genes of Wnt/β-catenin pathway were also analyzed by comparative PCR to determine the possible involvement. Results: In MSC-induced neuron-like cells, MET-associated transcription factors such as Snail, Slug, ZEB1, ZEB2, and Twist were significantly attenuated in expression level. The Mesenchymal marker Vimentin expression level was increased. Membrane protein E-cad was slightly down-regulated, while N-cad level was marginally elevated. Percentage of proliferating cells(S phase in cell cycle) markedly shrank from 40.42% for MSCs to 6.76% for MSC-derived neuron. Additionally, Wnt/β-catenin target genes β-catenin and c-myc were decreasingly expressed. Conclusion: Chemically induced trans-differentiation from MSC to neuron caused similar MET-featured alteration in gene expression and proliferation to known MET, which might be underlied by deactivation of Wnt/ β-catenin pathway.展开更多
Objective:To study the correlation of Wnt/β-catenin pathway changes with cytokines and apoptosis genes in diabetic foot wound.Methods:diabetic foot patients admitted to our hospital between February 2015 and October ...Objective:To study the correlation of Wnt/β-catenin pathway changes with cytokines and apoptosis genes in diabetic foot wound.Methods:diabetic foot patients admitted to our hospital between February 2015 and October 2017 were selected as the diabetic foot group,and non-diabetic patients admitted to our hospital during the same period were selected as the control group.Wound tissues of the diabetic foot group and normal skin tissues of the control group were collected to determine the mRNA expression levels of Wnt/β-catenin pathway molecules and apoptosis genes as well as the contents of cytokines.Results:The mRNA expression levels of Wnt1,β-catenin and B-cell lymphoma-2(Bcl-2)as well as the contents of vascular endothelial growth factor(VEGF),insulin-like growth factor-1(IGF-1)and basic fibroblast growth factor(bFGF)in the wounds of the diabetic foot group were significantly lower than those of the control group whereas the mRNA expression levels of glycogen synthase kinase-3(GSK-3β),Bcl-2-associated X protein(Bax)and cysteinyl aspartate specific proteinase-3(Caspase-3)as well as the contents of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were significantly higher than those of the control group;Wnt1 andβ-catenin were positively correlated with VEGF,IGF-1,bFGF and Bcl-2,and negatively correlated with TNF-α,IL-6,Bax and Caspase-3;GSK-3βwas negatively correlated with VEGF,IGF-1,bFGF,and Bcl-2,and positively correlated with TNF-α,IL-6,Bax and Caspase-3.Conclusion:Wnt/β-catenin pathway is inhibited in diabetic foot wound and it is related to the changes in cytokine secretion and apoptosis gene expression.展开更多
[Objectives]To explore the effects and mechanism of miR-15a-5p on oxaliplatin resistance in colorectal cancer HCT116/L cells.[Methods]The expression of miR-15a-5p in colorectal cancer sensitive cells HCT116 and resist...[Objectives]To explore the effects and mechanism of miR-15a-5p on oxaliplatin resistance in colorectal cancer HCT116/L cells.[Methods]The expression of miR-15a-5p in colorectal cancer sensitive cells HCT116 and resistant cells HCT116/L was detected by RT-qPCR method;the effect of oxaliplatin on the proliferation of HCT116 and HCT116/L cells was detected by MTT,and its IC_(50) and drug resistance fold of HCT116/L cells were calculated;the expressions of Wnt3a,β-Catenin and P-gp in HCT116 and HCT116/L cells were detected by Western Blot method.HCT-116/L cells were divided into 5 groups:blank control group HCT116/L,control group 1 transfected with miR-15a-5p mimics NC,experimental group 1 transfected with miR-15a-5p mimics,control group 2 transfected with miR-15a-5p inhibitor NC,and experimental group 2 transfected with miR-15a-5p inhibitor.The expression of miR-15a-5p in each group was detected by RT-qPCR method and the transfection efficiency was detected.The effect of different concentrations of oxaliplatin on the proliferation of cells in each group after transfection was detected by MTT and the half inhibitory concentration(50%inhibiting concentration,IC_(50))was calculated.The expressions of Wnt3a,β-catenin and P-gp in each group after transfection were detected by Western Blot method,and the expressions of Wnt3a,β-catenin and MDR1 mRNA in each group after transfection were detected by RT-qPCR method.[Results](i)RT-qPCR results showed that the expression of miR-15a-5p in HCT116/L cells was(0.16±0.05)significantly lower than that in HCT116 cells(P<0.05).(ii)The IC50 of oxaliplatin for HCT-116 cells and HCT116/L cells detected by MTT method were(13.51±2.62)and(103.08±12.29)μg/mL,respectively.The drug resistance index of HCT116/L was 7.63.(iii)Western Blot results showed that the expressions of Wnt3a,β-catenin and P-gp in HCT116/L cells were significantly higher than those in HCT-116 cells(P<0.05).(iv)After successful transfection,the IC50 of miR-15a-5p mimics group to oxaliplatin decreased to(40.78±2.47)μg/mL by MTT method,and its sensitivity to the drug was significantly improved compared with the control group.Western Blot results showed that the relative expressions of P-gp,Wnt3a andβ-catenin were significantly down-regulated(all P<0.05);RT-qPCR results showed that the relative expressions of MDR1,Wnt3a andβ-catenin mRNA were significantly down-regulated(all P<0.05).(v)After successful transfection,the expression of miR-15a-5p in the miR-15a-5p inhibitor transfection group was(0.38±0.04);MTT results showed that its IC50 for oxaliplatin was up-regulated to(132.77±7.97)μg/mL,and its sensitivity to chemotherapy drugs was significantly lower than that of the control group;Western Blot results showed that the relative expressions of P-gp,Wnt3a andβ-catenin were significantly up-regulated(all P<0.05);RT-qPCR results showed that the relative expressions of MDR1,Wnt3a andβ-catenin mRNA were significantly up-regulated(all P<0.05).[Conclusions]Up-regulation of miR-15a-5p expression can reverse the resistance of HCT116/L cell line to oxaliplatin.Up-regulation or down-regulation of miR-15a-5p will affect the expression of P-gp and Wnt/β-catenin signaling pathway-related proteins,suggesting that the mechanism of miR-15a-5p reversal of drug resistance may be related to the inhibition of Wnt/β-catenin pathway,thereby down-regulating the expression of P-gp.展开更多
基金National Nature Science Foundation of China(Nos.81960118,81860115,81760116 and 82060116)Guizhou Science and Technology Project:Qiankehe Foundation(No.(2020)1Y300)+8 种基金Natural Science Foundation of Sichuan(No.2022NSFSC0837)Science and Technology Project of Chengdu(No.2022-YF05-01811-SN)Science and Technology Project of Guizhou Province(No.YQK(2023)032)Guizhou Medical University Doctoral Start-Up Fund(No.gyfybsky-2021-27)Guizhou Medical University Doctoral Start-Up Fund(No.gyfybsky-2021-26)Guizhou Science and Technology Department(No.(2019)1259)Guizhou Science and Technology Department Guizhou Science and Technology Platform Talents(No.(2017)5718)Science and Technology Fund of Guizhou Provincial Health Commission(No.gzwki2021-382)The Affiliated Hospital of Guizhou Medical University Excellent Reserve Talent in 2023(No.gyfyxkrc-2023-06).
文摘Hepatocellular carcinoma(HCC),a common malignancy worldwide,still lacks effective clinical treatment.The study aimed to investigate the oncogenes that affect the progression of HCC and their possible mechanisms.In our study,we initially confirmed a higher level of PRDX2 in the bile of HCC patients compared to those with choledocholithiasis by 2-DE,LC-MS,and ELISA.Subsequently,we demonstrated the high expression of peroxiredoxin 2(PRDX2)in HCC based on the TCGA database and clinical sample analysis.Furthermore,PRDX2 overexpression enhanced the viability of HCC cells.And PRDX2 silencing induced senescence of HCC cells.In vivo,knockdown of PRDX2 significantly reduced the weight of xenograft tumors.PRDX2 also was found to activate the Wnt/β-catenin pathway by inducingβ-catenin nuclear translocation.Consequently,we proved that silencing PRDX2 could inhibit proliferation and Wnt/β-catenin pathway while promoting senescence in HCC cells.
基金The Fund of National Cancer Center Research and Development(26-A-4),The Grants-in-Aid for Scientific Research(Grant Nos.15K10451,16K10866 and 16K20063)from Japan Society for the Promotion of Science.
文摘An important factor in the emergence and progre sion of osteosarcoma(OS)is the dysregulated expression of microRNAs(miRNAs).Transcription factor 7-like 1(TCF7LI),a member of the T cell factor/lymphoid enhancer factor(TCF/LEF)transcription factor family,interacts with the Wnt signaling pathway regulator β-catenin and acts as a DNA-specific binding protein.This study sought to elucidate the impact of the interaction between miR 3293p and TCF7L1 on.the growth and apoptosis of OS and analyze the regulatory expression relationship between miRNA and mRNA in osteosarcoma cells using a variety of approaches.MiR329-3p was significantly downregulated,while TCF7L1 was considerably up-regulated in all examined OS cell lines.Additionally,a clinical comparison study was performed using the TCGA database.Subsequently,the regulatory relationship between miR-329-3p and TCF7L1 on the proliferation and apoptosis of OS cells was verified through in vitro and in vivo experiments.When miR 329-3p was transfected into the OS cell line,the expression of TCF7L1 decreased,the proliferation of OS cells was inhibited,the cytoskeleton disintegrated,and the nucleus condensed to fom apoptotic bodies.The expression of proteins that indicate apoptosis increased simultaneously.The cell cycle was arrested in the G0/G1 phase,and the G1/S transition was blocked.The introduction of miR 3293p also inhibited downstream Cyclin D1 of the Wnt pathway.Xenograf experiments indicated that the overexpression of miR-329-3p signi ficanly inhibited the growth of OS xenografts in nude mice,and the expression of TCF7L1 and C-Myc in tumor tssues decreased.MiR 329-3p was significantly reduced in OS cells and played a suppressive role in tumorigenesis and proliferation by targeting TCF7L1 both in vitro and in vivo.Osteosarcoma cell cycle arrest and pathway inhibition were observed upon the regulation of TCF7LI by miR 3293p.Summarizing these results,it can be inferred that miR.3293p exerts anticancer efects in osteosarcoma by inhibiting TCF7L1.
基金support from the Public Platform of Medical Research Center,Academy of Chinese Medical Science,Zhejiang Chinese Medical Universitysponsored by the National Natural Science Foundation of China(81973534,U1505226)。
文摘Increasing the osteogenic differentiation ability and decreasing the adipogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs)is a potential strategy for the treatment of osteoporosis(OP).Naturally derived oligosaccharides have shown significant anti-osteoporotic effects.Nystose(NST),an oligosaccharide,was isolated from the roots of Morinda officinalis How.(MO).The aim of the present study was to investigate the effects of NST on bone loss in ovariectomized mice,and explore the underlying mechanism of NST in promoting differentiation of BMSCs to osteoblasts.Administration of NST(40,80 and 160 mg/kg)and the positive control of estradiol valerate(0.2 mg/kg)for 8 weeks significantly prevented bone loss induced by ovariectomy(OVX),increased the bone mass density(BMD),improved the bone microarchitecture and reduced urine calcium and deoxypyridinoline(DPD)in ovariectomized mice,while inhibited the increase of body weight without significantly affecting the uterus weight.Furthermore,we found that NST increased osteogenic differentiation,inhibited adipogenic differentiation of BMSCs in vitro,and upregulated the expression of the key proteins of BMP and Wnt/β-catenin pathways.In addition,Noggin and Dickkopf-related protein-1(DKK-1)reversed the effect of NST on osteogenic differentiation and expression of the key proteins in BMP and Wnt/β-catenin pathway.The luciferase activities and the molecular docking analysis further supported the mechanism of NST.In conclusion,these results indicating that NST can be clinically used as a potential alternative medicine for the prevention and treatment of postmenopausal osteoporosis.
基金funded by the National Natural Science Foundation of China(Project No.81973615,No.81803910).
文摘Background:Actinidia chinensis Planch.roots(AcRoots)have been applied as an anti-inflammatory and antitumor drug in the treatment of gastric cancer(GC).However,their mechanisms against GC cells remain unclear.To investigate the anticancer effect of AcRoots in GC and the possible underlying mechanism by using network pharmacology.Methods:Differentially expressed genes between gastric precancerous lesions and cancer were analyzed in Gene Expression Omnibus datasets,and these genes were overlapped with potential targets of AcRoots.Potential targets and pathways for AcRoots treatment of GC predicted by network pharmacology.Furthermore,we used the GC cell line HGC27 to explore the molecular mechanisms in the context of hub genes in apoptosis,invasion,metastasis,and epithelial to mesenchymal transition-promoting factors.Molecular docking between hub targets and active drug components was also performed.Results:Network pharmacological analysis suggested that the potential mechanism was related to the Wnt pathway and predicted nine hub genes.In in vitro studies,AcRoots significantly decreased HGC27 cell viability and promoted apoptosis by upregulating caspase3 and downregulating Bcl2.Moreover,it suppressed invasion and metastasis as well as the expression of epithelial to mesenchymal transition-related factors.In addition,AcRoots affected the phosphorylation level of GSK3β(Ser9)in the Wnt pathway to promote the degradation ofβ-catenin,resulting in the downregulation of the downstream target genes c-myc,cyclin D1 and snail.All the experimental results were consistent with the network pharmacology results.Conclusion:This study combined network pharmacology with in vitro experiments to provide valid evidence for the clinical promotion of AcRoots.
文摘BACKGROUND Exosomal miRNAs play crucial roles in many central nervous system diseases.Cerebral small vessel disease(CVSD)is a small vessel disease that is affected by various factors.This study aimed to investigate the role of exosomal miR-320e in the Wnt/β-catenin pathway stimulated by oxidative stress and assess its clinical correlation with psychiatric symptoms in patients with CVSD.AIM To explore whether exosomal miR-320e could suppress the Wnt/β-catenin pathway and play a protective role in CVSD progression,as well as examine its potential correlation with cognitive impairment and depression in patients with CVSD.METHODS Differentially expressed exosomal miRNAs were filtered by sequencing plasma exosomes from patients with CVSD and healthy controls.Bioinformatics and dual luciferase analyses were used to confirm the binding of miR-320e to Wnt2,and the mRNA and protein levels of downstream components in the Wnt/β-catenin pathway were evaluated when overexpressed or with knockdown of miR-320e under H2O2-induced oxidative stress.In addition,Wnt2-targeting siRNA was used to confirm the role of miR-320e in the Wnt2-mediated inhibition of the Wnt/β-catenin pathway.A retrospective analysis was conducted among patients with CVSD to confirm the correlation between miR-320e expression and the severity of cognitive impairment and depression,which were quantified using the Montreal Cognitive Assessment(MoCA)/Executive Function Assessment(EFA),and the Hamilton Depression Scale(HAMD)/Beck Depression Inventory(BDI),respectively.RESULTS High-throughput sequencing revealed that exosomal miR-320e was downregulated in patients with CVSD.Bioinformatics analysis and dual-luciferase reporter gene experiments showed that exosomal miR-320e inhibited the Wnt/β-catenin pathway in response to oxidative stress by targeting the 3'noncoding region of Wnt2.Uptake of exosomes carrying miR-320e into endothelial cells could also target Wnt2 and inhibit the Wnt2/β-catenin pathway.Elevated miR-320e expression may protect patients with CVSD from relatively severe cognitive impairment and depression,as it was found to have a positive correlation with the MoCA/EFA and HAMD/BDI scores.CONCLUSION Our results suggest that exosomal miR-320e suppresses the Wnt/β-catenin pathway and may play a protective role in CVSD progression.
基金a Grant-in-Aid for Scientific Research from National Natural Science Foundation of China,No. 81860120 and 81860104Guangxi Natural Science Foundation,No. 2017GXNSFBA198134, 2017GXNSFAA198299 and 2015GXNSFCA139024
文摘BACKGROUND Hepatic fibrosis is a common pathological process of chronic liver diseases with various causes,which can progress to cirrhosis.AIM To evaluate the effect and mechanism of action annexin(Anx)A1 in liver fibrosis and how this could be targeted therapeutically.METHODS CCl4(20%)and active N-terminal peptide of AnxA1(Ac2-26)and N-formylpeptide receptor antagonist N-Boc-Phe-Leu-Phe-Leu-Phe(Boc2)were injected intraperitoneally to induce liver fibrosis in eight wild-type mice/Anxa1 knockout mice,and to detect expression of inflammatory factors,collagen deposition,and the role of the Wnt/β-catenin pathway in hepatic fibrosis.RESULTS Compared with the control group,AnxA1,transforming growth factor(TGF)-β1,interleukin(IL)-1βand IL-6 expression in the liver of mice with hepatic fibrosis induced by CCl4 was significantly increased,which promoted collagen deposition and expression ofα-smooth muscle actin(α-SMA),collagen type I and connective tissue growth factor(CTGF),and increased progressively with time.CCl4 induced an increase in TGF-β1,IL-1βand IL-6 in liver tissue of AnxA1 knockout mice,and the degree of liver inflammation and fibrosis and expression ofα-SMA,collagen I and CTGF were significantly increased compared with in wild-type mice.After treatment with Ac2-26,expression of liver inflammatory factors,degree of collagen deposition and expression of a-SMA,collagen I and CTGF were decreased compared with before treatment.Boc2 inhibited the anti-inflammatory and antifibrotic effects of Ac2-26.AnxA1 downregulated expression of the Wnt/β-catenin pathway in CCl4-induced hepatic fibrosis.In vitro,lipopolysaccharide(LPS)induced hepatocyte and hepatic stellate cell(HSC)expression of AnxA1.Ac2-26 inhibited LPS-induced RAW264.7 cell activation and HSC proliferation,decreased expression ofα-SMA,collagen I and CTGF in HSCs,and inhibited expression of the Wnt/β-catenin pathway after HSC activation.These therapeutic effects were inhibited by Boc2.CONCLUSION AnxA1 inhibited liver fibrosis in mice,and its mechanism may be related to inhibition of HSC Wnt/β-catenin pathway activation by targeting formylpeptide receptors to regulate macrophage function.
基金Supported by Henan Provincial Natural Science Foundation of China,No.212300410242Youth Project Jointly Constructed by Henan Provincial Health Commission and the Ministry,No.SBGJ202103008Henan Young and Middle-aged Health Science and Technology Innovation Excellent Youth Talent Training Project of China,No.YXKC2021047.
文摘BACKGROUND Mesenchymal stem cells(MSCs)have been applied to treat degenerative articular diseases,and stromal cell-derived factor-1α(SDF-1α)may enhance their therapeutic efficacy.However,the regulatory effects of SDF-1αon cartilage differentiation remain largely unknown.Identifying the specific regulatory effects of SDF-1αon MSCs will provide a useful target for the treatment of degenerative articular diseases.AIM To explore the role and mechanism of SDF-1αin cartilage differentiation of MSCs and primary chondrocytes.METHODS The expression level of C-X-C chemokine receptor 4(CXCR4)in MSCs was assessed by immunofluorescence.MSCs treated with SDF-1αwere stained for alkaline phosphatase(ALP)and with Alcian blue to observe differentiation.Western blot analysis was used to examine the expression of SRY-box transcription factor 9,aggrecan,collagen II,runt-related transcription factor 2,collagen X,and matrix metalloproteinase(MMP)13 in untreated MSCs,of aggrecan,collagen II,collagen X,and MMP13 in SDF-1α-treated primary chondrocytes,of glycogen synthase kinase 3β(GSK3β)p-GSK3βandβ-catenin expression in SDF-1α-treated MSCs,and of aggrecan,collagen X,and MMP13 in SDF-1α-treated MSCs in the presence or absence of ICG-001(SDF-1αinhibitor).RESULTS Immunofluorescence showed CXCR4 expression in the membranes of MSCs.ALP stain was intensified in MSCs treated with SDF-1αfor 14 d.The SDF-1αtreatment promoted expression of collagen X and MMP13 during cartilage differentiation,whereas it had no effect on the expression of collagen II or aggrecan nor on the formation of cartilage matrix in MSCs.Further,those SDF-1α-mediated effects on MSCs were validated in primary chondrocytes.SDF-1αpromoted the expression of p-GSK3βandβ-catenin in MSCs.And,finally,inhibition of this pathway by ICG-001(5μmol/L)neutralized the SDF-1α-mediated up-regulation of collagen X and MMP13 expression in MSCs.CONCLUSION SDF-1αmay promote hypertrophic cartilage differentiation in MSCs by activating the Wnt/β-catenin pathway.These findings provide further evidence for the use of MSCs and SDF-1αin the treatment of cartilage degeneration and osteoarthritis.
基金the National Natural Science Foundation of China(Nos.81703973,81973669,81703912,and 81603484)National Key R&D Program of China(No.2018YFC1705600)+4 种基金Research Projects of Key Disease for Practice Development of National TCM Clinical Research Bases(No.JDZX2015172)Science and Technology Research Program of Hubei Education Department for Young Investigators(No.Q20172003)The Green Seedlings Program of Hubei University of Chinese Medicine(No.HUCM[2016]224)Special Program for Advanced Researchers of Hubei University of Chinese Medicine(No.5114-100724)Li Han-min National Famous Old Chinese Medicine Experts Inheritance Studio.
文摘The activation of the Wnt/β-catenin signaling pathway in oval cells after liver injury is implicated in hepatocarcinogenesis.Diwu Yanggan capsule is a Chinese herbal medicine that has been used for treating liver disorder.The present study aimed to examine the mechanism by which Diwu Yanggan inhibits liver carcinogenesis,and the involvement of the Wnt/β-catenin signaling pathway.Diwu Yanggan capsule was administered to 2-acetaminofluorene/partial hepatectomy(2-AAF/PH)rats,a murine model of liver injury.The biomarkers of oval cells and key proteins in the Wnt/p-catenin signaling pathway were assessed on postoperative day 8,10,14,17,19 and 22.The results showed that treatment with Diwu Yanggan was associated with reduced expression of oval cell and stem cell biomarkers in the 2-AAF/PH animals.The expression pattern of key proteins in the Wnt/β-catenin pathway was altered in Diwu Yanggan-treated animals,indicating that the Diwu Yanggan treatment accelerated the activation of the Wnt/β-catenin pathway in the initial stage and contributed to its deactivation in the later stage.Histological findings indicated that hepatocyte proliferation was suppressed in Diwu Yanggan-treated animals,compared with untreated 2-AAF/PH animals.Taken together,Diwu Yanggan capsule may reduce the risk of hepatocarcinogenesis by modulating the Wnt/β-catenin signaling pathway.
文摘The neuropsychiatric disease named obsessive-compulsive disorder is composed by obsessions and/or compulsions.Obsessive-compulsive disorder etiologies are undefined.However,numerous mechanisms in several localizations are implicated.Some studies showed that both glutamate,inflammatory factors and oxidative stress could have main functions in obsessive-compulsive disorder.Glycogen synthase kinase-3β,the major negative controller of the WNT/β-catenin pathway is upregulated in obsessive-compulsive disorder.In obsessive-compulsive disorder,some studies presented the actions of the different circadian clock genes.WNT/β-catenin pathway and circadian clock genes appear to be intricate.Thus,this review focuses on the interaction between circadian clock genes and the WNT/β-catenin pathway in obsessive-compulsive disorder.
基金This study was supported by the National Key Research and Development Program of China(No.2018YFC0311203)the Key Research and Development Plan in Shandong Province(No.2016YYSP017).
文摘In this study,fucoidans were extracted from the sea cucumber Thelenota ananas(Ta-FUCs)by enzymatic degradation.Four products with molecular weights of 1380.3,524.0,182.4,and 110.3 kDa were obtained,and the Ta-FUC showing optimal anti-adipogenic activities was determined.Results of MTT and Oil red O staining analyses showed that the Ta-FUCs inhibited the proliferation and differentiation of 3T3-L1 adipocytes.Futhermore,Ta-FUCs significantly downregulated the key transcriptional factors,such as SREBP-1c,PPARγ,and C/EBPαof adipocytes.The Ta-FUCs also activated Wnt/β-catenin pathway-related genes,such asβ-catenin,LRP5,and FrZ.The Ta-FUCs suppressed lipid accumulation in 3T3-L1 adipocytes possibly by decreasing the expression of genes ACC,FAS,ME,GPAT,DGAT,and PILN,which are important in the synthesis of fatty acids and triglycerides;and by increasing the expression of genes PPARα,CPT-1α,and ACOX,which are crucial in fatty acidβ-oxidation.The anti-adipogenic activities initially increased and then declined with decreasing molecular weight.Among the Ta-FUCs,the 182.4 kDa Ta-FUC exhibited optimal bioactivities.This study reports for the first time that Ta-FUCs can prevent obesity by regulating the differentiation and lipid accumulation of adipocytes.
基金This study was financially supported by grants from the National Natural Science Foundation of China(82071119,82071142,81700938,81772876,81800942).
文摘Osteoporosis is a frequently occurring bone remodeling disorder worldwide with one characteristic being decreasing bone mineral density and a predisposition to bone fracture,which diminishes patients’quality of life.Several studies showed that imbalance between the osteogenesis and adipogenesis of bone marrow mesenchymal stem cells(BMSCs)took part in the development of osteoporosis.In previous study,we found MIR22HG regulated the osteogenesis of human BMSCs positively.In this study,we found that MIR22HG was decreased during the adipogenesis of human BMSCs and exerted negative effects on adipogenesis with the involvement of Wnt/β-catenin signaling pathway both in vitro and in vivo.Nitazoxanide could inhibit Wnt signaling and relieve MIR22HG’s suppression on adipogenesis.These findings indicated that MIR22HG had great potential in clinical application for osteoporosis treatment and prevention.
文摘Recent studies showed that activation of Wnt/β-catenin pathway promoted the differentiation of osteoblast-like cells in the arterial calcification, but its mechanism remains unknown. In this study, the hypothesis that Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by upregulating the expression of receptor activator of NF-κB ligand (RANKL) was examined. LiCl was used to activate the Wnt/β-catenin pathway. The differentiation of osteoblast-like cells was observed by Von Kossa staining, calcium content assay, alkaline phosphatase (ALP) activity assay, and detection of osteocalcin expression. Real-time PCR was performed to detect the expression of RANKL and osteoprotegerin (OPG, the decoy receptor of RANKL) during the osteoblast-like cell differentiation. Different concentrations of OPG were added to the culture media respectively to inhibit the function of RANKL, and the change in the differentiation of osteoblast-like cells was evaluated. The results showed that when the Wnt/β-catenin pathway was activated by LiCl, the expression of RANKL was significantly in-creased, which coincided with the differentiation of osteoblast-like cells (P<0.05), and the OPG treatment could partly attenuate the promoting effect of Wnt/β-catenin pathway on the differentiation of osteoblast-like cells (P<0.05), but it failed to completely abolish such effect. It was concluded that activation of Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by both RANKL-dependent and RANKL-independent mechanisms.
文摘In this study,the hypothesis that Wnt/β-catenin pathway is involved in the arterial calcification by regulating the osteoprotegerin(OPG)/receptor activator of NF-κB ligand(RANKL)system was tested.Theβ-catenin expression was measured in the warfarin-induced calcified arteries and the osteoblast-like cells differentiating from smooth muscle cells(SMCs)by immunohistochemistry and Western blotting.The Wnt/β-catenin pathway was activated or inhibited by lithium chloride(LiCl)or dickkopf 1(DKK1)in vitro and in vivo.Then the calcification level was determined by von Kossa staining,Ca^2+content assay,and alkaline phosphatase(ALP)activity assay.The expression levels of osteocalcin,OPG and RANKL were detected by Western blotting or real-time PCR.The results showed that in calcified arteries and OBL cells,the activation of Wnt/β-catenin pathway significantly enhanced the calcification as evidenced by increased von Kossa stains,Ca^2+contcnts,ALP activities,and osteocalcin expression levels(P<0.05),and it promoted the RANKL expression(P<0.05),but slightly affected the OPG expression.These results indicated that the activation of Wnt/β-catenin pathway worsens the arterial calcification,probably by promoting the RANKL expression.
文摘Gastric cancer remains one of the most common cancers worldwide and one of the leading cause for cancerrelated deaths. Gastric adenocarcinoma is a multifactorial disease that is genetically, cytologically and architecturally more heterogeneous than other gastrointestinal carcinomas.The aberrant activation of the Wnt/β-catenin signaling pathway is involved in the development and progression of a significant proportion of gastric cancer cases. This review focuses on the participation of the Wnt/b-catenin pathway in gastric cancer by offering an analysis of the relevant literature published in this field. Indeed, it is discussed the role of key factors in Wnt/β-catenin signaling and their downstream effectors regulating processes involved in tumor initiation, tumor growth, metastasis and resistance to therapy. Available data indicate that constitutive Wnt signalling resulting from Helicobacter pylori infection and inactivation of Wnt inhibitors(mainly by inactivating mutations and promoter hypermethylation) play an important role in gastric cancer. Moreover, a number of recent studies confirmed CTNNB1 and APC as driver genes in gastric cancer. The identification of specific membrane, intracellular, and extracellular components of the Wnt pathway has revealed potential targets for gastric cancer therapy. High-throughput "omics" approaches will help in the search for Wnt pathway antagonist in the near future.
基金Zhejiang Provincial Natural Science Foundation of China,No.LQ18H160011 and No.LY20H030011.
文摘BACKGROUND Our previous study demonstrated that RBBP4 was upregulated in colon cancer and correlated with poor prognosis of colon cancer and hepatic metastasis.However,the potential biological function of RBBP4 in colon cancer is still unknown.AIM To investigate the biological role and the potential mechanisms of RBBP4 in colon cancer progression.METHODS Real-time polymerase chain reaction and western blot analysis were used to detect the expression of RBBP4 in colon cancer cell lines.The cell proliferation and viability of SW620 and HCT116 cells with RBBP4 knockdown was detected by Cell Counting Kit-8 and 5-ethynyl-2’-deoxyuridine staining.The transwell assay was used to detect the invasion and migration capabilities of colon cancer cells with RBBP4 knockdown.Flow cytometry apoptosis assay was used to detect the apoptosis of colon cancer cells.Western blotting analysis was used to detect the expression of epithelial-mesenchymal transition and apoptosis related markers in colon cancer.The nuclear translocation ofβ-catenin was examined by Western blotting analysis in colon cancer cells with RBBP4 knockdown.The TOPFlash luciferase assay was used to detect the effect of RBBP4 on Wnt/β-catenin activation.The rescue experiments were performed in colon cancer cells treated with Wnt/β-catenin activator LiCl and RBBP4 knockdown.RESULTS We found that RBBP4 was highly expressed in colon cancer cell lines.The 5-ethynyl-2’-deoxyuridine assay showed that knockdown of RBBP4 significantly inhibited cell proliferation.RBBP4 inhibition reduced cell invasion and migration via regulating proteins related to epithelial-mesenchymal transition.Knockdown of RBBP4 significantly inhibited survivin-mediated apoptosis.Mechanistically,the TOPFlash assay showed that RBBP4 knockdown increased activity of the Wnt/β-catenin pathway.Meanwhile,RBBP4 knockdown suppressed nuclear translocation ofβ-catenin.With Wnt/β-catenin activator,rescue experiments suggested that the role of RBBP4 in colon cancer progression was dependent on Wnt/β-catenin pathway.CONCLUSION RBBP4 promotes colon cancer development via increasing activity of the Wnt/β-catenin pathway.RBBP4 may serve as a novel therapeutic target in colon cancer.
基金Supported by Agencia Nacional de Promoción Científica y Tecnológica,No. PICT-2013-1441Consejo Nacional de Investigaciones Científicas y Técnicas,No. PIP11220150100350+4 种基金Instituto Nacional del Cáncer Asistencia Financiera ⅡRESOL 493/14, No. 2002-4395-14-1Instituto Nacional del Cáncer Asistencia Financiera Ⅲ-2016-2017, RESOL-2016-1006-E-APN-MS,No. 2002-3862-16-1Universidad Nacional del Sur (PGI)Argentina,No. 24/B230 and No. 24/B303
文摘Colorectal cancer(CRC)continues to be one of the main causes of death from cancer because patients progress unfavorably due to resistance to current therapies.Dysregulation of the Wnt/β-catenin pathway plays a fundamental role in the genesis and progression of several types of cancer,including CRC.In many subtypes of CRC,hyperactivation of theβ-catenin pathway is associated with mutations of the adenomatous polyposis coli gene.However,it can also be associated with other causes.In recent years,studies of the tumor microenvironment(TME)have demonstrated its importance in the development and progression of CRC.In this tumor nest,several cell types,structures,and biomolecules interact with neoplastic cells to pave the way for the spread of the disease.Cross-communications between tumor cells and the TME are then established primarily through paracrine factors,which trigger the activation of numerous signaling pathways.Crucial advances in the field of oncology have been made in the last decade.This Minireview aims to actualize what is known about the central role of the Wnt/β-catenin pathway in CRC chemoresistance and aggressiveness,focusing on crosscommunication between CRC cells and the TME.Through this analysis,our main objective was to increase the understanding of this complex disease considering a more global context.Since many treatments for advanced CRC fail due to mechanisms involving chemoresistance,the data here exposed and analyzed are of great interest for the development of novel and effective therapies.
文摘Objective: To investigate the effect of arthroscopic debridement combined with intra-articular sodium hyaluronate injection on the Wnt/β-catenin pathway in joint fluid of patients with knee osteoarthritis. Methods: Patients undergoing arthroscopic debridement for knee osteoarthritis in the hospital between June 2014 and April 2017 were selected as the research subjects and randomly divided into the observation group who accepted arthroscopic debridement combined with intra-articular sodium hyaluronate injection and the control group who accepted arthroscopic debridement alone. The Wnt/β-catenin pathway molecules as well as downstream collagen metabolism molecules and bone metabolism molecules in the joint fluid were measured before surgery and 7 d after surgery. Results: 7 d after surgery, Wnt1, Wnt5a, Wnt7b, β-catenin, MMP1, MMP13, CTX-I, CTX-II, ADAMTS4, ADAMTS5, RANK, RANKL and TRACP5b mRNA expression in joint fluid of both groups of patients were higher than those before surgery whereas OPG and OC contents were lower than those before surgery, and Wnt1, Wnt5a, Wnt7b, β-catenin, MMP1, MMP13, CTX-I, CTX-II, ADAMTS4, ADAMTS5, RANK, RANKL and TRACP5b mRNA expression in joint fluid of observation group were significantly lower than those of control group whereas OPG and OC contents were significantly higher than those of control group. Conclusion: Arthroscopic debridement combined with intra-articular sodium hyaluronate injection can inhibit the collagen metabolism and bone metabolism disorder mediated by Wnt/β-catenin after surgery.
文摘Objective: To observe MET-associated alteration during the trans-differentiation from MSCs to neuron-like cells, and to explore the possible molecular mechanism. Methods: Bone marrow MSCs were isolated from rat femur and purified in continuous cell culture. After induced differentiation to neuron-like cells by the combination of butylated hydroxyanisole(BHA)and dimethyl sulfoxide(DMSO), cells were tested by comparative polymerase chain reaction(PCR) for the relative expression of MET biomarkers and transcription factors, and for cell cycle by flow cytometry. Meanwhile, target genes of Wnt/β-catenin pathway were also analyzed by comparative PCR to determine the possible involvement. Results: In MSC-induced neuron-like cells, MET-associated transcription factors such as Snail, Slug, ZEB1, ZEB2, and Twist were significantly attenuated in expression level. The Mesenchymal marker Vimentin expression level was increased. Membrane protein E-cad was slightly down-regulated, while N-cad level was marginally elevated. Percentage of proliferating cells(S phase in cell cycle) markedly shrank from 40.42% for MSCs to 6.76% for MSC-derived neuron. Additionally, Wnt/β-catenin target genes β-catenin and c-myc were decreasingly expressed. Conclusion: Chemically induced trans-differentiation from MSC to neuron caused similar MET-featured alteration in gene expression and proliferation to known MET, which might be underlied by deactivation of Wnt/ β-catenin pathway.
基金Health and Family Planning Commission of Hubei Province(No:WJ2017M183).
文摘Objective:To study the correlation of Wnt/β-catenin pathway changes with cytokines and apoptosis genes in diabetic foot wound.Methods:diabetic foot patients admitted to our hospital between February 2015 and October 2017 were selected as the diabetic foot group,and non-diabetic patients admitted to our hospital during the same period were selected as the control group.Wound tissues of the diabetic foot group and normal skin tissues of the control group were collected to determine the mRNA expression levels of Wnt/β-catenin pathway molecules and apoptosis genes as well as the contents of cytokines.Results:The mRNA expression levels of Wnt1,β-catenin and B-cell lymphoma-2(Bcl-2)as well as the contents of vascular endothelial growth factor(VEGF),insulin-like growth factor-1(IGF-1)and basic fibroblast growth factor(bFGF)in the wounds of the diabetic foot group were significantly lower than those of the control group whereas the mRNA expression levels of glycogen synthase kinase-3(GSK-3β),Bcl-2-associated X protein(Bax)and cysteinyl aspartate specific proteinase-3(Caspase-3)as well as the contents of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were significantly higher than those of the control group;Wnt1 andβ-catenin were positively correlated with VEGF,IGF-1,bFGF and Bcl-2,and negatively correlated with TNF-α,IL-6,Bax and Caspase-3;GSK-3βwas negatively correlated with VEGF,IGF-1,bFGF,and Bcl-2,and positively correlated with TNF-α,IL-6,Bax and Caspase-3.Conclusion:Wnt/β-catenin pathway is inhibited in diabetic foot wound and it is related to the changes in cytokine secretion and apoptosis gene expression.
文摘[Objectives]To explore the effects and mechanism of miR-15a-5p on oxaliplatin resistance in colorectal cancer HCT116/L cells.[Methods]The expression of miR-15a-5p in colorectal cancer sensitive cells HCT116 and resistant cells HCT116/L was detected by RT-qPCR method;the effect of oxaliplatin on the proliferation of HCT116 and HCT116/L cells was detected by MTT,and its IC_(50) and drug resistance fold of HCT116/L cells were calculated;the expressions of Wnt3a,β-Catenin and P-gp in HCT116 and HCT116/L cells were detected by Western Blot method.HCT-116/L cells were divided into 5 groups:blank control group HCT116/L,control group 1 transfected with miR-15a-5p mimics NC,experimental group 1 transfected with miR-15a-5p mimics,control group 2 transfected with miR-15a-5p inhibitor NC,and experimental group 2 transfected with miR-15a-5p inhibitor.The expression of miR-15a-5p in each group was detected by RT-qPCR method and the transfection efficiency was detected.The effect of different concentrations of oxaliplatin on the proliferation of cells in each group after transfection was detected by MTT and the half inhibitory concentration(50%inhibiting concentration,IC_(50))was calculated.The expressions of Wnt3a,β-catenin and P-gp in each group after transfection were detected by Western Blot method,and the expressions of Wnt3a,β-catenin and MDR1 mRNA in each group after transfection were detected by RT-qPCR method.[Results](i)RT-qPCR results showed that the expression of miR-15a-5p in HCT116/L cells was(0.16±0.05)significantly lower than that in HCT116 cells(P<0.05).(ii)The IC50 of oxaliplatin for HCT-116 cells and HCT116/L cells detected by MTT method were(13.51±2.62)and(103.08±12.29)μg/mL,respectively.The drug resistance index of HCT116/L was 7.63.(iii)Western Blot results showed that the expressions of Wnt3a,β-catenin and P-gp in HCT116/L cells were significantly higher than those in HCT-116 cells(P<0.05).(iv)After successful transfection,the IC50 of miR-15a-5p mimics group to oxaliplatin decreased to(40.78±2.47)μg/mL by MTT method,and its sensitivity to the drug was significantly improved compared with the control group.Western Blot results showed that the relative expressions of P-gp,Wnt3a andβ-catenin were significantly down-regulated(all P<0.05);RT-qPCR results showed that the relative expressions of MDR1,Wnt3a andβ-catenin mRNA were significantly down-regulated(all P<0.05).(v)After successful transfection,the expression of miR-15a-5p in the miR-15a-5p inhibitor transfection group was(0.38±0.04);MTT results showed that its IC50 for oxaliplatin was up-regulated to(132.77±7.97)μg/mL,and its sensitivity to chemotherapy drugs was significantly lower than that of the control group;Western Blot results showed that the relative expressions of P-gp,Wnt3a andβ-catenin were significantly up-regulated(all P<0.05);RT-qPCR results showed that the relative expressions of MDR1,Wnt3a andβ-catenin mRNA were significantly up-regulated(all P<0.05).[Conclusions]Up-regulation of miR-15a-5p expression can reverse the resistance of HCT116/L cell line to oxaliplatin.Up-regulation or down-regulation of miR-15a-5p will affect the expression of P-gp and Wnt/β-catenin signaling pathway-related proteins,suggesting that the mechanism of miR-15a-5p reversal of drug resistance may be related to the inhibition of Wnt/β-catenin pathway,thereby down-regulating the expression of P-gp.
