Effects Of MicroRNA-1271-5p On The Biological Behavior Of Hepatocellular Carcinoma And Its Radiosensitivity

Objective:This study aims to investigate the role of mi R-1271-5p in tumors and LIHC.Alsoaims to determine whether mi R-1271-5p inhibits cell proliferation and enhances the radiosensitivity by targeting cyclin-dependent kinase 1(CDK1)in hepatocellular carcinoma(HCC).Investigate the role of AVIL in cell proliferation,invasion,migration,and Wnt/?-catenin/EMT in hepatocellular carcinoma(HCC).MATERIALS AND METHODS:The correlation between mi RNA-1271 and LIHC was analyzed by bioinformatics.The expression level of mi R-1271-5p in cells was examined using quantitative reverse transcription polymerase chain reaction(RT-q PCR).Western blot analysis was performed to detect the expression level of CDK1 in HCC cell lines.Bioinformatics analysis was used to predict some potential target genes of mi R-1271-5p.The Dual-Luciferase Reporter Assay system was utilized to determine the direct relationship between mi R-1271-5p and CDK1.SMMC-7721 cells were overexpressed for mi R-1271-5p to obtain a stable Lv-mi R-1271-5p cell line.Cell proliferation,radiosensitivity,and cell cycle were assessed by Cell Counting Kit 8(CCK8)assay,flow cytometry,and clone formation assay.We first predicted the expression of AVIL from the Cancer Genome Atlas(TCGA),and then detected the expression level of AVIL in different HCC cell lines by q-PCR.Subsequently,AVIL was silenced and a series of experiments were performed to determine the effects of AVIL on HCC cells proliferation,migration,invasion and Wnt /?-catenin / EMT in vitro and in vivo.RESULTS : The expression of mi R-1271-5p is reduced in many tumors,especially LIHC.The role of mi R-1271-5p in the development of LIHC may be related to the regulation of the core protein FKBP1 A in the corresponding PPI network of mi R-1271-5p.Its expression levels in the HCC cell lines were significantly lower than those in normal human liver cell line.Bioinformatics analysis indicated CDK1 was a potential target of mi R-1271-5p.Dual-Luciferase Reporter Assay confirmed that CDK1 is a direct target gene of mi R-1271-5p.With overexpression of mi R-1271-5p in SMMC-7721 and Hu H-7cells,cell proliferation was decreased,radiosensitivity was enhanced,cell cycle distribution was altered,and the growth of transplanted tumors in nude mice was significantly reduced.Mi R-1271-5p overexpression enhanced radiosensitivity,which could be reduced by CDK1 overexpression.The prediction results of TCGA and the cell experiments showed that the expression of AVIL in HCC were significantly higher than normal.Silencing AVIL had significantly decreased the AVIL expression,the proliferation,and the clonogenicity in HCC cells,while the apoptosis rate was significantly increased.In vivo,silencing of AVIL in SMMC-7721 cells also can inhibited tumor growth.Moreover,the migration and invasion capacity of silencing AVIL cells was decreased,in vivo studies also indicated that the silencing AVIL reduced pulmonary migration of HCC cells.Furthermore,silencing AVIL not only can up-regulates E-cadherin in HCC cells,xenografts and lung metastases,while advillin,?-catenin,N-cadherin,and C-myc are down-regulated.But also effectively inhibite both Wnt /?-catenin / EMT in HCC cells in vitro and in vivo.CONCLUTIONS:Overall,our findings suggested that mi R-1271-5p plays an important role in the occurrence and development of tumors,especially LIHC.mi R-1271-5p inhibits cell proliferation and enhances the radiosensitivity of HCC cell lines by targeting CDK1.Silencing AVIL can decreases proliferation,invasion,migration,and Wnt/?-catenin/EMT in HCC both in vitro and in vivo.