Effects And Underlying Mechanisms Of Tβ10 On Cell Invasion,migration And Apoptosis Of Hepatocellular Carcinoma

Objective: In this study,bioinformatics methods were used to analyze the expression level of Tβ10 in HCC data sets and its correlation with the survival and prognosis of HCC patients,to investigate the expression level of Tβ10 in human hepatocellular carcinoma cell lines,and the effect and possible mechanism of Tβ10’s down-regulation on the migration,invasion and apoptosis of human hepatocellular carcinoma cell lines,in order to find biomarkers for diagnosis and prognosis evaluation,and potential therapeutic targets of hepatocellular carcinoma.Methods:1.The expression level of Tβ10 in HCC tissues was analyzed in Oncomine database,and then its clinical value was further analyzed in GEPIA database.2.RT-PCR detected Tβ10 mRNA expression levels of two HCC cell lines including HepG2 and MHCC-97L and normal cell line QSG-7701.The HCC cells with down-regulation of Tβ10 were conduced by siRNA transient transfection technique.Transwell invasion assay and wound healing assay were performed to measure the effect of Tβ10 on cell invasion and migration.Cell apoptosis was measured with combined staining of PI and DAPI.In addition,the Bcl-2/Bax expression ratio and the EMT molecular markers including E-cadherin and N-cadherin were determined by Western blot.Results:1.Bioinformatics analysis results showed that Tβ10 overexpressed in most hepatocellular carcinoma data sets in Oncomine database,and its high expression warned poor clinical prognosis,while it was pointless for clinical stage of HCC in GEPIA database.2.Compared with human normal liver cells,the expression level of Tβ10 mRNA was increased in HepG2 and MHCC-97L cell lines(P<0.05).Tβ10 mRNA expression was down-regulated in HepG2 and MHCC-97L cell lines after transfection with Tβ10siRNA(P<0.05),and the ability of migration and invasion was decreased(P<0.05).The results of DAPI/PI staining showed that the cell nucleus in the experimental group were condensed and concentrated,showing bright blue fluorescence,or were lobulated and fragmented,which performanced significantly more than those in the negative control group.Western blot results showed that inhibition of Tβ10 significantly down-regulated the expression of N-cadherin in MHCC-97L and HepG2(P<0.05),and up-regulated the expression of E-cadherin.Bcl-2/Bax expression ratio was also up-regulated(P<0.05).Conclusion: Tβ10 overexpresses in HCC cells and HCC tissues,compared with the human normal liver cells and the adjacent or normal liver tissues.High expression of Tβ10 is associated with poor clinical prognosis.Tβ10 may promotes the invasion and migration of HCC cells via epithelial mesenchymal transformation.Tβ10 can also inhibit the apoptosis of HCC cells,the mechanism of apoptosis may be related to the down-regulation of Bcl-2/Bax expression ratio.Therefore,Tβ10 is expected to be a biomarker for the diagnosis and prognostic evaluation of HCC,and provides a new potential target for the biological therapy of HCC patients.