Cinobufotalin prevents bone loss induced by ovariectomy in mice through the BMPs/SMAD and Wnt/β-catenin signaling pathways

Background:Osteoporosis is a chronic bone disease characterized by bone loss and decreased bone strength.However,current anti-resorptive drugs carry a risk of various complications.The deep learning-based efficacy prediction system(DLEPS)is a forecasting tool that can effectively compete in drug screening and prediction based on gene expression changes.This study aimed to explore the protective effect and potential mechanisms of cinobufotalin(CB),a traditional Chinese medicine(TCM),on bone loss.Methods:DLEPS was employed for screening anti-osteoporotic agents according to gene profile changes in primary osteoporosis.Micro-CT,histological and morphological analysis were applied for the bone protective detection of CB,and the osteogenic differentiation/function in human bone marrow mesenchymal stem cells(hBMMSCs)were also investigated.The underlying mechanism was verified using qRT-PCR,Western blot(WB),immunofluorescence(IF),etc.Results:A safe concentration(0.25mg/kg in vivo,0.05μM in vitro)of CB could effectively preserve bone mass in estrogen deficiency-induced bone loss and promote osteogenic differentiation/function of hBMMSCs.Both BMPs/SMAD and Wnt/β-catenin signaling pathways participated in CB-induced osteogenic differentiation,further regulating the expression of osteogenesis-associated factors,and ultimately promoting osteogenesis.Conclusion:Our study demonstrated that CB could significantly reverse estrogen deficiency-induced bone loss,further promoting osteogenic differentiation/function of hBMMSCs,with BMPs/SMAD and Wnt/β-catenin signaling pathways involved.

Pachymic acid exerts antitumor activities by modulating the Wnt/β-catenin signaling pathway via targeting PTP1B

Objective:To determine the inhibitory effects of pachymic acid on lung adenocarcinoma(LUAD)cells and elucidate its underlying mechanism.Methods:CCK-8,wound healing,Transwell,Western blot,tube formation,and immunofluorescence assays were carried out to measure the effects of various concentrations of pachymic acid on LUAD cell proliferation,metastasis,angiogenesis as well as autophagy.Subsequently,molecular docking technology was used to detect the potential targeted binding association between pachymic acid and protein tyrosine phosphatase 1B(PTP1B).Moreover,PTP1B was overexpressed in A549 cells to detect the specific mechanisms of pachymic acid.Results:Pachymic acid suppressed LUAD cell viability,metastasis as well as angiogenesis while inducing cell autophagy.It also targeted PTP1B and lowered PTP1B expression.However,PTP1B overexpression reversed the effects of pachymic acid on metastasis,angiogenesis,and autophagy as well as the expression of Wnt3a andβ-catenin in LUAD cells.Conclusions:Pachymic acid inhibits metastasis and angiogenesis,and promotes autophagy in LUAD cells by modulating the Wnt/β-catenin signaling pathway via targeting PTP1B.

Effects of Helicobacter pylori and Moluodan on the Wnt/β-catenin signaling pathway in mice with precancerous gastric cancer lesions

BACKGROUND Helicobacter pylori(H.pylori)is the primary risk factor for gastric cancer(GC),the Wnt/β-Catenin signaling pathway is closely linked to tumourigenesis.GC has a high mortality rate and treatment cost,and there are no drugs to prevent the progression of gastric precancerous lesions to GC.Therefore,it is necessary to find a novel drug that is inexpensive and preventive to against GC.AIM To explore the effects of H.pylori and Moluodan on the Wnt/β-Catenin signaling pathway and precancerous lesions of GC(PLGC).METHODS Mice were divided into the control,N-methyl-N-nitrosourea(MNU),H.pylori+MNU,and Moluodan groups.We first created an H.pylori infection model in the H.pylori+MNU and Moluodan groups.A PLGC model was created in the remaining three groups except for the control group.Moluodan was fed to mice in the Moloudan group ad libitum.The general condition of mice were observed during the whole experiment period.Gastric tissues of mice were grossly and microscopically examined.Through quantitative real-time PCR(qRT-PCR)and Western blotting analysis,the expression of relevant genes were detected.RESULTS Mice in the H.pylori+MNU group showed the worst performance in general condition,gastric tissue visual and microscopic observation,followed by the MNU group,Moluodan group and the control group.QRT-PCR and Western blotting analysis were used to detect the expression of relevant genes,the results showed that the H.pylori+MNU group had the highest expression,followed by the MNU group,Moluodan group and the control group.CONCLUSION H.pylori can activate the Wnt/β-catenin signaling pathway,thereby facilitating the development and progression of PLGC.Moluodan suppressed the activation of the Wnt/β-catenin signaling pathway,thereby decreasing the progression of PLGC.

ZNF554 Inhibits Endometrial Cancer Progression via Regulating RBM5 and Inactivating WNT/β-Catenin Signaling Pathway

Objective:Uterine corpus endometrial carcinoma(UCEC),a kind of gynecologic malignancy,poses a significant risk to women’s health.The precise mechanism underlying the development of UCEC remains elusive.Zinc finger protein 554(ZNF554),a member of the Krüppel-associated box domain zinc finger protein superfamily,was reported to be dysregulated in various illnesses,including malignant tumors.This study aimed to examine the involvement of ZNF554 in the development of UCEC.Methods:The expression of ZNF554 in UCEC tissues and cell lines were examined by qRT-PCR and Western blot assay.Cells with stably overexpressed or knocked-down ZNF554 were established through lentivirus infection.CCK-8,wound healing,and Transwell invasion assays were employed to assess cell proliferation,migration,and invasion.Propidium iodide(PI)staining combined with fluorescence-activated cell sorting(FACS)flow cytometer was utilized to detect cell cycle distribution.qRT-PCR and Western blotting were conducted to examine relative mRNA and protein levels.Chromatin immunoprecipitation assay and luciferase reporter assay were used to explore the regulatory role of ZNF554 in RNA binding motif 5(RBM5).Results:The expression of ZNF554 was found to be reduced in both UCEC samples and cell lines.Decreased expression of ZNF554 was associated with higher tumor stage,decreased overall survival,and reduced disease-free survival in UCEC.ZNF554 overexpression suppressed cell proliferation,migration,and invasion,while also inducing cell cycle arrest.In contrast,a decrease in ZNF554 expression resulted in the opposite effect.Mechanistically,ZNF554 transcriptionally regulated RBM5,leading to the deactivation of the Wingless(WNT)/β-catenin signaling pathway.Moreover,the findings from rescue studies demonstrated that the inhibition of RBM5 negated the impact of ZNF554 overexpression onβ-catenin and p-glycogen synthase kinase-3β(p-GSK-3β).Similarly,the deliberate activation of RBM5 reduced the increase inβ-catenin and p-GSK-3βcaused by the suppression of ZNF554.In vitro experiments showed that ZNF554 overexpression-induced decreases in cell proliferation and migration were counteracted by RBM5 knockdown.Additionally,when RBM5 was overexpressed,it hindered the improvements in cell proliferation and migration caused by reducing the ZNF554 levels.Conclusion:ZNF554 functions as a tumor suppressor in UCEC.Furthermore,ZNF554 regulates UCEC progression through the RBM5/WNT/β-catenin signaling pathway.ZNF554 shows a promise as both a prognostic biomarker and a therapeutic target for UCEC.

Effects of Qigongwan on Wnt/β-catenin Signaling Pathway in Rats with Polycystic Ovary syndrome

[Objectives] To explore the therapeutic effect and mechanism of Qigongwan on PCOS model rats by detecting the changes in sex hormone levels in rats with polycystic ovary syndrome (PCOS), and observing the effects of ovarian pathological morphological changes, apoptosis and expression of Wnt/β-β catenin signaling pathway protein. [Methods] Ten of 40 female SD rats were randomly selected as the normal group, and the other 30 rats were treated with letrozole combined with high-fat diet to establish the PCOS rat model. After successful modeling, the model group was randomly divided into Qigongwan group, positive Daying-35 (Ethinylestradiol and Cyproterone Acetate Tablets) group and model group, with 10 rats in each group. Qigongwan group was given 14.7 g/(kg·d) by gavage, Daying-35 group was given 0.21 mg/(kg·d) by oral gavage, and normal group and model group were given the same amount of distilled water, and the intervention lasted for 21 d. ELISA method was used to detect the levels of hormones such as follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), estradiol (E 2) and progesterone (P) in serum. HE staining was used to observe the pathological morphological changes of ovarian tissues;TUNEL staining was used to observe apoptosis of ovarian tissue granule cells;the expression of Wnt, β-catenin protein in rat ovarian tissue was detected by immunohistochemistry. [Results] (i) Compared with the model group, Qigongwan group and Daying-35 group could significantly increase serum E 2 and P levels, significantly reduce serum T levels ( P <0.01), significantly reduce serum LH levels and LH/FSH ratio ( P <0.01), and increase serum FSH levels ( P <0.05) in different degrees. (ii)The results of HE staining showed that compared with the model group, Qigongwan and Daying-35 groups could improve follicular development and reduce atretic follicles in different degrees. Compared with Daying-35 group, the number of GC layers in Qigongwan group was significantly increased. (iii) The results of TUNEL staining showed that compared with the model group, the rate of TUNEL-positive cells in the Qigongwan group and Daying-35 group decreased significantly ( P <0.01). (iv) The immunohistochemical results showed that compared with the model group, the expression levels of wnt and β-catenin in the Qigongwan group and the Daying-35 group increased in different degrees ( P <0.05), and the expression range increased. [Conclusions] Qigongwan can regulate the secretion level of sex hormones such as FSH and LH, improve the pathological damage of ovarian tissue, and inhibit apoptosis of ovarian granule cells, and its mechanism may be related to the activation of Wnt/β-catenin signaling pathway.

TCM Intervention on Wnt/β-Catenin signaling pathway in the treatment of diabetic nephropathy

Diabetic nephropathy(DN)is the most serious microvascular complication of diabetes mellitus,which is highly prevalent worldwide.Abnormal activation of Wnt/β-catenin signaling pathway is an important mechanism of renal damage induced by hyperglycemia.Many studies have shown that TCM has the advantages of high efficiency and safety in the prevention and treatment of DN.Some TCM monomers and compounds repair podocyte function and inhibit transdifferentiation process by inhibiting the activation of Wnt/β-catenin signaling pathway,thus playing a protective role in kidney.Based on this,this paper will review the existing research results and related mechanisms of TCM intervention in Wnt/β-catenin signaling pathway in the treatment of DN,in order to promote the more effective and reasonable application of TCM in clinical practice.

The Role and Mechanism of LncRNA-p21 in Regulating Gastric Cancer Metastasis by Mediating Wnt/β-Catenin Signaling Pathway

Objective:To study the mechanism of lncRNA p21 inhibiting the growth and metastasis of human gastric cancer SGC7901/GES-1 cells by mediating the Wnt/β-catenin signaling pathway.Methods:Lentiviral overexpression of lncRNA-p21 in human gastric cancer SGC7901/GES-1 cell transfections was observed and analyzed for in vitro migration,invasion,cell morphology and proliferation.Besides.Wnt/β-catenin signaling pathway was tested for direct involvement in lncRNA-p21-mediated inhibition of gastric cancer cell growth and proliferation.Wnt/β-catenin signaling pathway was validated using Li-C1.Results:Gastric cancer SGC7901/GES-1 cells in the overexpression of lncRNA-p21 showed changes in stellate morphology,low invasion,or spindle-shaped morphology.LncRNA-p21 inhibited the growth and proliferation of gastric cancer SGC7901/GES-1 cells both in vivo and in vitro,and Wnt/β-catenin signaling pathway mediated the proliferation,invasion,and migration of lncRNA-p21 on gastric cancer SGC7901/GES-1 cells.Conclusion:LncRNA-p21 can inhibit the growth and metastasis of gastric cancer SGC7901/GES-1 cells in vitro and in vivo,and the inhibition of lncRNA p21 is mainly mediated by inhibiting the Wnt/β-catenin signaling pathway.

Neuroprotective effect of rapamycin on spinal cord injury via activation of the Wnt/β-catenin signaling pathway

The Wnt/β-catenin signaling pathway plays a crucial role in neural development, axonal guid- ance, neuropathic pain remission and neuronal survival. In this study, we initially examined the effect of rapamycin on the Wnt/β-catenin signaling pathway after spinal cord iniury, by intraperitoneally injecting spinal cord injured rats with rapamycin over 2 days. Western blot analysis and immunofluorescence staining were used to detect the expression levels of β-catenin protein, caspase-3 protein and brain-derived neurotrophic factor protein, components of the Wnt/β-catenin signaling pathway. Rapamycin increased the levels of β-catenin and brain-derived neurotrophic factor in the injured spinal cord, improved the pathological morphology at the injury site, reduced the loss of motor neurons, and promoted motor functional recovery in rats after spinal cord injury. Our experimental fndings suggest that the neuroprotective effect of rapamycin intervention is mediated through activation of the Wnt/β-catenin signaling pathway after spinal cord injury.

MiR-19a-3p regulates the Forkhead box F2-mediated Wnt/β-catenin signaling pathway and affects the biological functions of colorectal cancer cells

BACKGROUND Colorectal cancer(CRC)is one of the most common malignancies worldwide.AIM To explore the expression of microRNA miR-19a-3p and Forkhead box F2(FOXF2)in patients with CRC and the relevant mechanisms.METHODS Sixty-two CRC patients admitted to the hospital were enrolled into the study group,and sixty healthy people from the same period were assigned to the control group.Elbow venous blood was sampled from the patients and healthy individuals,and blood serum was saved for later analysis.MiR-19a-3p mimics,miR-19a-3p inhibitor,miR-negative control,small interfering-FOXF2,and short hairpin-FOXF2 were transfected into HT29 and HCT116 cells.Then quantitative polymerase chain reaction was performed to quantify the expression of miR-19a-3p and FOXF2 in HT29 and HCT116 cells,and western blot(WB)analysis was conducted to evaluate the levels of FOXF2,glycogen synthase kinase 3 beta(GSK-3β),phosphorylated GSK-3β(p-GSK-3β),β-catenin,p-β-catenin,α-catenin,Ncadherin,E-cadherin,and vimentin.The MTT,Transwell,and wound healing assays were applied to analyze cell proliferation,invasion,and migration,respectively,and the dual luciferase reporter assay was used to determine the correlation of miR-19a-3p with FOXF2.RESULTS The patients showed high serum levels of miR-19a-3p and low levels of FOXF2,and the area under the curves of miR-19a-3p and FOXF2 were larger than 0.8.MiR-19a-3p and FOXF2 were related to sex,tumor size,age,tumor-nodemetastasis staging,lymph node metastasis,and differentiation of CRC patients.Silencing of miR-19a-3p and overexpression of FOXF2 suppressed the epithelialmesenchymal transition,invasion,migration,and proliferation of cells.WB analysis revealed that silencing of miR-19a-3p and FOXF2 overexpression significantly suppressed the expression of p-GSK-3β,β-catenin,N-cadherin,and vimentin;and increased the levels of GSK-3β,p-β-catenin,α-catenin,and Ecadherin.The dual luciferase reporter assay confirmed that there was a targeted correlation of miR-19a-3p with FOXF2.In addition,a rescue experiment revealed that there were no differences in cell proliferation,invasion,and migration in HT29 and HCT116 cells co-transfected with miR-19a-3p-mimics+sh-FOXF2 and miR-19a-3p-inhibitor+si-FOXF2 compared to the miR-negative control group.CONCLUSION Inhibiting miR-19a-3p expression can upregulate the FOXF2-mediated Wnt/β-catenin signaling pathway,thereby affecting the epithelial-mesenchymal transition,proliferation,invasion,and migration of cells.Thus,miR-19a-3p is likely to be a therapeutic target in CRC.

Expression analysis of eight amphioxus genes involved in the Wnt/β-catenin signaling pathway

The Wnt/β-catenin signaling pathway plays a crucial role in the embryonic development of metazoans. Although the pathway has been studied extensively in many model animals, its function in amphioxus, the most primitive chordate, remains largely uncharacterized. To obtain basic data for functional analysis, we identified and isolated seven genes （Lrp5/6, Dvl, APC, Ckla, CklS, Gsk3β, and Gro） of the Wnt/β-catenin signaling pathway from the amphioxus （Branchiostoma floridae） genome. Phylogenetic analysis revealed that amphioxus had fewer members of each gene family than that found in vertebrates. Whole-mount in situ hybridization showed that the genes were maternally expressed and broadly distributed throughout the whole embryo at the cleavage and blastula stages. Among them, Dvl was expressed asymmetrically towards the animal pole, while the others were evenly distributed in all blastomeres. At the mid-gastrula stage, the genes were specifically expressed in the primitive endomesoderm, but displayed different patterns. When the embryo developed into the neurula stage, the gene expressions were mainly detected in either paraxial somites or the tail bud. With the development of the embryo, the expression levels further decreased gradually and remained only in some pharyngeal regions or the tail bud at the larva stage. Our results suggest that the Wnt/β-catenin pathway might be involved in amphioxus somite formation and posterior growth, but not in endomesoderm specification.

Role of Wnt/β-catenin Signaling Pathway in the Mechanism of Calcification of Aortic Valve

Aortic valve calcification is a common disease in the elderly, but its cellular and molecular mechanisms are not clear. In order to verify the hypothesis that Wnt/β-catenin signaling pathway is involved in the process of calcification of aortic valve, porcine aortic valve interstitial cells（VICs） were isolated, cultured and stimulated with oxidized low density lipoprotein（ox-LDL） for 48 h to induce the differentiation of VICs into osteoblast-like cells. The key proteins and genes of Wnt/β-catenin signaling pathway, such as glycogen synthase kinase 3β（GSK-3β） and β-catenin, were detected by using Western blotting and real-time polymerase chain reaction（PCR）. The results showed that the VICs managed to differentiate into osteoblast-like cells after the stimulation with ox-LDL and the levels of proteins and genes of GSK-3β and β-catenin were increased significantly in VICs after stimulation for 48 h（P0.05）. It is suggested that Wnt/β-catenin signaling pathway may play a key role in the differentiation of VICs into osteoblast-like cells and make great contribution to aortic valve calcification.

Effects of Wnt/β-catenin signaling pathway on the proliferation and invasive growth of cells in mantle cell lymphoma

Objective:To study the effects of Wnt/β-catenin signaling pathway on the proliferation and invasive growth of cells in mantle cell lymphoma.Methods: Patients who were diagnosed with mantle cell lymphoma by lymph node biopsy in Tongji Hospital? Tongji Medical College Huazhong University of Science & Technology between March 2015 and May 2017 were selected as lymphoma group of the research, and patients who were diagnosed with reactive hyperplasia by lymph node biopsy during the same period were selected as the control group. The protein expression of Wnt/β-catenin signaling pathway molecules in lymph node tissues were determined by enzyme-linked immunosorbent assay kit, and the mRNA expression of proliferation genes and invasion genes were determined by fluorescence quantitative PCR kit.Results:β-catenin, p-GSK-3β and Tcf/Lef protein levels as well as C-myc, CyclinD1, Est-1, MMP9 and MMP26 mRNA expression in lymph node tissues of lymphoma group were significantly higher than those of control group whereas DKK1 and WIF-1 protein levels as well as Bax, Apaf1, Caspase-3, TNFAIP3, TIMP2 and TIMP4 mRNA expression were significantly lower than those of control group. C-myc, CyclinD1, Est-1, MMP9 and MMP26 mRNA expression in lymphoma tissues were positively correlated withβ-catenin, p-GSK-3β and Tcf/Lef protein levels, and negatively correlated with DKK1 and WIF-1 protein levels;Bax, Apaf1, Caspase-3, TNFAIP3, TIMP2 and TIMP4 mRNA expression were negatively correlated withβ-catenin, p-GSK-3β and Tcf/Lef protein levels, and positively correlated with DKK1 and WIF-1 protein levels.Conclusion: The activation of Wnt/β-catenin signaling pathway can promote the proliferation and invasive growth of cells in mantle cell lymphoma.

Mechanism of miR-21 via Wnt/β-catenin signaling pathway in human A549 lung cancer cells and Lewis lung carcinoma in mice

Objective:To study the mechanism of effect of miR-21 via Wnt/ β-catenin signaling pathway in human A549 lung cancer cells and Lewis lung carcinoma in mice.Methods:The effect of miR-21 on A549 cells were detected by MTT method.MiR-21 expression levels were overexpressed or inhibited in A549 cells by transfecting with miR-21 mimics or inhibitors.Correlation among key molecules(Wnt1,β-catenin.CyclinD1 and miR-21) of mRNA and protein levels in Wnt/β-catenin signaling pathway were studied by Real-time PCR and Western blot hybridization assay.Invasive ability of A549 cells was determined via Transwell chamber cell invasion assay;the role of miR-21 in A549 cells was explored via the Wnt/β-catenin signaling pathway.A Lewis lung carcinoma animal model was established to detect miR-21 expressions in tumor animals and controlled animal tissues,and verify expression changes of the above moleculesin the Wnt / β-catenin signaling pathway was determined in the animal level.Results:MTT assay results showed that miR-21 overexpression could markedly enhance cell absorbance value;that is,miR-21 could increase the ability proliferation of A549 cells.β-catenin and CyclinD1 expression levels were significantly higher in miR-21 mimic transfected cells(P<0.05),and Wnt 1 gene had no significant change.Wnt 1,β-catenin and CyclinD1 gene expression showed no significant change when miR-21 expression was suppressed,compared with controls.After cells were transfected with miR-21 mimics,cell invasion assay revealed that the perforated cells was significantly higher than the perforated cells in the control group(P<0.01).Lewis lung assay revealed that miR-21 expression levels in the Lewis lung carcinoma were significantly higher;and at the same time.Wnt1,β-catenin and CyclinD1 gene expression levels were significantly increased,compared to controls.Conclusions:In A549 human lung cancer cells and Lewis lung carcinoma in mice,key molecules β-catenin and CyclinD1 of miR-21 expressions and the Wnt/ β-catenin signaling pathway are positively correlated.

Effects of Er-Zhi-Wan on Microarchitecture and Regulation of Wnt/β-catenin Signaling Pathway in Alveolar Bone of Ovariectomized Rats

Recent studies have shown that Er-Zhi-Wan（EZW）, a traditional Chinese medicine consisting of Herba Ecliptae（HE） and Fructus Ligustri Lucidi（FLL）, had a definite antiosteoporotic effect on osteoporotic femur, but its effect on osteoporosis of alveolar bone remains unknown. In the present study, we investigated the effects of Er-Zhi-Wan（EZW） on the microarchitecture and the regulation of Wnt/β-catenin signaling pathway in the alveolar bone of ovariectomized rats. Thirty Sprague-Dawley rats were randomly divided into three groups： sham operation group（sham, n=10）, ovariectomy（OVX） group（n=10）, and OVX with EZW treatment group（EZW group, n=10）. From one week after ovariectomy, EZW（100 mg/mL） or vehicle（distilled water） was fed（1 mL/100 g） once per day for 12 weeks until the sacrifice of the rats. The body weights were measured weekly. After sacrifice, the sera and mandible were collected and routinely prepared for the measurement of alveolar trabecular microarchitecture, serum levels of E2, bone-specific alkaline phosphatase（BALP） and tartrate-resistant acid phosphatase 5b（TRAP5b）, as well as mandibular mRNA expression of Wnt/β-catenin signaling pathway molecules wnt3a, low-density lipoprotein receptor-related protein 5（LRP5）, β-catenin and dickkopf homolog 1（DKK1）. The results showed that EZW treatment significantly prevented the body weight gain, degradation of alveolar trabecular microarchitecture and alveolar bone loss in the OVX rats. Furthermore, we observed that EZW could increase the serum levels of E2 and BALP, and decrease levels of serum TRAP5b in EZW group compared with vehicle group. In addition, RT-PCR results revealed that EZW upregulated the expression levels of wnt3a, LRP5 and β-catenin, and reduced the expression of DKK1 in OVX rats. Taken together, our results suggested that EZW may have potential anti-osteoporotic effects on osteoporotic alveolar bone by stimulating Wnt/LRP5/β-catenin signaling pathway.

The Wnt/β-catenin signaling pathway in the tumor microenvironment of hepatocellular carcinoma

The Wnt/β-catenin signaling pathway regulates many aspects of tumor biology,and many studies have focused on the role of this signaling pathway in tumor cells.However,it is now clear that tumor development and metastasis depend on the two-way interaction between cancer cells and their environment,thereby forming a tumor microenvironment(TME).In this review,we discuss how Wnt/β-catenin signaling regulates cross-interactions among different components of the TME,including immune cells,stem cells,tumor vasculature,and noncellular components of the TME in hepatocellular carcinoma.We also investigate their preclinical and clinical insights for primary liver cancer intervention,and explore the significance of using Wnt/β-catenin mutations as a biomarker to predict resistance in immunotherapy.

Roles of Wnt/β-catenin signaling pathway related microRNAs in esophageal cancer

MicroRNAs(mi RNAs)are endogenous,noncoding,single-stranded small RNAs that regulate expression of tumor suppressor genes and oncogenes and are involved in almost all tumor-related processes.Mi RNA dysregulation plays an important role in the occurrence and development of esophageal cancer through specific signal pathways,including the Wnt/β-catenin signaling pathway,and is closely related to the malignant characteristics of esophageal cancer.The interaction between mi RNAs and the Wnt/β-catenin signaling pathway,which is specifically expressed in esophageal cancer tissues,shows potential as a new biomarker and therapeutic target.This article reviews the role of mi RNAs related to the Wnt pathway in the carcinogenesis of esophageal carcinoma and its role in Wnt signal transduction.The content of this review can be used as the basis for formulating or improving the treatment strategy of esophageal cancer.

Effect of miR-124 Overexpression under Wnt/β-catenin Signaling Pathway on Inflammatory Factor and Epidermal Growth Factor in Rats with Precancerous Gastric Lesions

Objective:To investigate the effects of miR-124 overexpression on inflammatory factors and epidermal growth factor in rats with gastric precancerous lesions by modulating the Wnt/β-catenin signaling pathway.Methods:Eighty SPF-grade Wistar rats were selected as the research subjects.After adaptive feeding,they were divided into 4 groups using a random number table method.The control group was fed a regular diet,while the model group,low expression group,and overexpression group of rats were treated with N-methyl-N’-nitro-N-nitrosoguanidine to establish a model of gastric cancer precancerous lesions.After successful modeling,the control group and model group rats were injected intraperitoneally with 0.9%sodium chloride solution,the low-expression group rats were injected intraperitoneally with miR-124-5p inhibitor,and the over-expression group rats were injected intraperitoneally with miR-124-5p mimic.After 8 weeks of continuous injection,the levels of miR-124,Wnt/β-catenin signaling pathway-related proteins,inflammatory factors,and epidermal growth factor were observed and compared among the four groups of rats.Results:The differences in the relative expression of miR-124 among the four groups of rats were statistically significant(P<0.05);the differences in the expression of Wnt protein andβ-catenin protein among the four groups of rats were statistically significant(P<0.05),and the expression of Wnt protein andβ-catenin protein in the overexpression group was significantly higher than that of the control group,and significantly lower than that in the model group and the low-expression group;the levels of IL-1β,IL-6,and TNF-αof rats in the four groups of rats IL-1β,IL-6 and TNF-αlevels were statistically different(P<0.05),and the levels of IL-1β,IL-6 and TNF-αin rats in the overexpression group were higher than those in the model group and significantly lower than those in the model group and the low-expression group(P<0.05);the differences in the levels of EGF,HER1 and HER2 among the four groups of rats were statistically different(P<0.05),and the levels of EGF levels were higher than those of the control group,and EGF and HER1 were significantly lower than those of the model and low expression groups(P<0.05).Conclusion:Mi R-124 overexpression helps to reduce gastric mucosal damage and inflammatory response in rats with gastric precancerous lesions,and its mechanism of action may be linked to the inhibition of the Wnt/β-catenin signaling pathway.

Xiao Tan San Jie Fang Hampers the Growth of Colon Cancer Stem Cells through the Wnt/β-catenin Signaling Pathway

Objective: The purpose of this research was to examine the potential anticancer properties of Xiao Tan San Jie Fang(XTSJF) and its potential mechanism of action against colorectal cancer. Materials and Methods: HCT116 cells were induced into HCT116 spheres in DMEM/F12 medium by treatment with epidermal growth factor + fibroblast growth factor + leukemia inhibitory factor + B27. The proliferation ability and stemness of HCT116 spheres was examined. Various concentrations of XTSJF were used to treat HCT116 spheres to observe the impact on proliferation, apoptosis, and expression of stem cell markers. Next, Wnt/β-catenin pathway-related factor proteins were detected. Results: The findings revealed that XTSJF suppressed cell growth and induced cell death in HCT116 cells in a dosage-dependent manner. Similarly, XTSJF promotes apoptosis, inhibits cell proliferation, prolongs survival, and maintains the expression of stem cells through the Wnt/-catenin/TCF4 axis. XTSJF also inhibits AKT activity and subsequently activates glycogen synthesis kinase-3β expression, inhibiting Wnt/beta-catenin pathway activity and downstream target gene transcript expression. The Wnt/β-catenin signaling pathway is inhibited by the XTSJF, leading to the suppression of colon cancer stem cell proliferation. Conclusion: Xiaotan Sanjie Prescription inhibited colon cancer stem cell growth through Wnt/β-catenin signaling pathway.

Efficacy of Yisui granule(益髓颗粒)on myelodysplastic syndromes in SKM-1 mouse xenograft model through suppressing Wnt/β-catenin signaling pathway

OBJECTIVE:To unmask the underlying mechanisms of Yisui granule(益髓颗粒,YSG)for the treatment of Myelodysplastic syndromes(MDS).METHODS:Our study used an SKM-1 mouse xenograft model of MDS to explore the anti-tumor potential of YSG and its safety,assess its effect on overall survival(OS),and evaluate whether its mechanism is associated with the demethylation of the secreted frizzled related protein 5(s FRP5)gene and suppressing Wnt/β-catenin pathway.Bisulfite amplicon sequencing was applied to detect the level of methylation of the s FRP5 gene;western blotting,immunofluorescence staining,and real-time Polymerase Chain Reaction were performed to detect DNA methyltransferase 1(DNMT1),s FRP5,and other Wnt/β-catenin pathway-related m RNA and protein expression.RESULTS:The results showed that high-dosage YSG exerted an anti-tumor effect similar to that of decitabine,improved OS,and reduced long-term adverse effects in the long term.Mechanically,YSG reduced the expression of DNMT1 methyltransferase,decreased the methylation,and increased the expression of the Wnt/β-catenin pathway antagonist-s FRP5.Furthermore,components of the Wnt/β-catenin pathway,including Wnt3a,β-catenin,c-Myc,and cyclin D1,were down-regulated in response to YSG,suggesting that YSG could treat MDS by demethylating the s FRP5 gene and suppressing the Wnt/β-catenin pathway.CONCLUSIONS:Our findings demonstrated that YSG could be used alone or in combination with decitabine to improve outcomes in the MDS animal model,providing an alternative solution for treating MDS.

Complement factor Ⅰ knockdown inhibits colon cancer development by affecting Wnt/β-catenin/c-Myc signaling pathway and glycolysis

BACKGROUND Colon cancer(CC)occurrence and progression are considerably influenced by the tumor microenvironment.However,the exact underlying regulatory mechanisms remain unclear.AIM To investigate immune infiltration-related differentially expressed genes(DEGs)in CC and specifically explored the role and potential molecular mechanisms of complement factor I(CFI).METHODS Immune infiltration-associated DEGs were screened for CC using bioinformatics.Quantitative reverse transcription polymerase chain reaction was used to examine hub DEGs expression in the CC cell lines.Stable CFI-knockdown HT29 and HCT116 cell lines were constructed,and the diverse roles of CFI in vitro were assessed using CCK-8,5-ethynyl-2’-deoxyuridine,wound healing,and transwell assays.Hematoxylin and eosin staining and immunohistochemistry staining were employed to evaluate the influence of CFI on the tumorigenesis of CC xenograft models constructed using BALB/c male nude mice.Key proteins associated with glycolysis and the Wnt pathway were measured using western blotting.RESULTS Six key immune infiltration-related DEGs were screened,among which the expression of CFI,complement factor B,lymphoid enhancer binding factor 1,and SRY-related high-mobility-group box 4 was upregulated,whereas that of fatty acid-binding protein 1,and bone morphogenic protein-2 was downregulated.Furthermore,CFI could be used as a diagnostic biomarker for CC.Functionally,CFI silencing inhibited CC cell proliferation,migration,invasion,and tumor growth.Mechanistically,CFI knockdown downregulated the expression of key glycolysis-related proteins(glucose transporter type 1,hexokinase 2,lactate dehydrogenase A,and pyruvate kinase M2)and the Wnt pathway-related proteins(β-catenin and c-Myc).Further investigation indicated that CFI knockdown inhibited glycolysis in CC by blocking the Wnt/β-catenin/c-Myc pathway.CONCLUSION The findings of the present study demonstrate that CFI plays a crucial role in CC development by influencing glycolysis and the Wnt/β-catenin/c-Myc pathway,indicating that it could serve as a promising target for therapeutic intervention in CC.