MicroRNA-361-5p Suppresses The Tumorigenesis Of Hepatocellular Carcinoma Through Targeting WT1 And Suppressing WNT/β-catenin Pathway

Hepatocellular carcinoma(HCC)is the third most common malignant tumor after gastric cancer and esophageal malignant tumor.Moreover,the early symptoms of HCC are not obvious.The advanced HCC is mainly characterized by liver pain,angular,jaundice,ascites and other symptoms.However,the etiology and exact molecular mechanism of HCC are not fully understood until now.It is currently believed that the pathogenesis of HCC is a complex process of multiple factors and steps,affected by its environment and genetics.Although the tumorigenesis of HCC can be seen in all age groups,it is relatively rare in children.Most Chinese HCC patients are between 45 and 65 years old.However,clinical studies have found that the incidence of young people is significantly increased in recent years.The incidence of HCC in men is higher than that in women,and the ratio of male to female is about 3:1.Surgery,radiotherapy and chemotherapy combined with traditional Chinese medicine are widely used in clinical practice.However,the cure rate of patients with advanced HCC is still low due to cell proliferation.The one-year survival rate after HCC resection is higher than 80%,and the five-year survival rate is over 50%.In addition,the prognosis of HCC patients was found to be correlated with clinical and pathological type.Therefore,the biomarkers for early detection,early diagnosis and early treatment of HCC should be developed.Previous studies have shown that microRNAs(miRNAs)play important roles in the pathogenesis of human cancers by affecting expressions of target genes.Moreover,many miRNAs that are oncogenes and tumor suppressors have been found to be involved in tumorigenesis of HCC.For example,miR-199 functioned as a suppressive miRNA by downregulating Regulators of G-protein signaling 17 in HCC.In contrast,miR-219-5p promoted tumor growth and cell metastasis in HCC by regulating Cadherin 1.In recent years,miR-361-5p has been found to exert different effect in some human cancers.Wu et al proposed that miR-361-5p facilitated the development of cervical cancer by mediating epithelial-mesenchymal transition(EMT).In contrast,the inhibitory effect of miR-361-5p was observed in prostate cancer.Moreover,miR-361-5p suppressed chemoresistance by modulating PI3K/AKT/mTOR pathway in gastric cancer.However,the specific role of miR-361-5p is still unclear in HCC by now.Therefore,we designed this research to explore its function in HCC.Wilms’ tumor-1(WT1)has been found to play a diagnostic and prognostic role in high-grade serous ovarian carcinoma.Moreover,abnormal expression of WT1 had been identified in human breast cancer.And WT1 was found be upregulated in desmoids tumors.Liu et al reported that high WT1 expression was associated with aggressive clinical features of ovarian cancer.Besides that,the WNT/β-cadherin pathway has been demonstrated to regulate the development of HCC.For example,the carcinogenic effect of miR-324-3p was found in HCC through activating the WNT/β-catenin pathway.Cui et al proposed that miR-337 regulated PI3K/AKT and WNT/β-catenin signaling pathways to inhibit HCC progression.However,the interaction between miR-361-5p and WT1 or WNT/β-catenin signaling pathway has not been investigated in HCC.In the current study,the potential function of miR-361-5p was explored in HCC.Moreover,downstream regulatory mechanism of miR-361-5p was also investigated in HCC.The interaction between miR-361-5p and WT1 or WNT/β-catenin signaling pathway was verified in this study.Objective:MicroRNA-361-5p(miR-361-5p)has been found to be involved in the pathogenesis of several human cancers.However,the specific role of miR-361-5p is still unclear in hepatocellular carcinoma(HCC).Therefore,this study was designed to elucidate the function of miR-361-5p in HCC.Method:The expression levels of miR-361-5p and Wilms’ tumor-1(WT1)were detected by quantitative real-time polymerase chain reaction(qRT-PCR)assay.Moreover,the function of miR-361-5p was examined through Cell Counting Kit-8(CCK-8)and Transwell assays.Protein expressions were examined via Western Blot analysis and Immunocyto chemical assay.Tumor growth of HCC was observed via xenograft tumor formation assay.The relationship between miR-361-5p and WT1 was verified by dual luciferase assay.Results:Downregulation of miR-361-5p was identified in HCC,which predicted a worse prognosis in HCC patients.Furthermore,it was found that miR-361-5p suppressed cell proliferation,migration and invasion in HCC by inhibiting WT1.MiR-361-5p also inhibited tumor growth of HCC.Besides that,miR-361-5p suppressed EMT and negatively activated the WNT/a-catenin pathway in HCC.Conclusion:MiR-361-5p suppressed tumorigenesis of HCC by targeting WT1 and inactivating WNT/β-catenin pathway.