The Mechanism Of MiR-93Induced Epithelial Mesenchymal Transition Via EphA4to Regulate Me Recurrence Of Hepatocellular Carcinoma Following Liver Transplantation

From the angle of miRNomics, we investigated the expression patterns of miRNomics in normal liver tissues and HCCs to find the differential expression of miRNA which are involved in the progressions of HCC through Illumina Solexa MPSS. We found that microRNA-93(miR-93) is overexpressed in HCCs than those in normal liver tissues. Emerging evidence suggests that miRNAs play a vital role in the carcinogenesis and progression of HCC, however the underlying mechanism of miR-93in HCC is still unclear. Therefore, a better understanding of the molecular mechanisms underlying HCC recurrence, identifying more accurate predictive biomarkers and establishing a novel model that can assess tumor recurrence and prognosis is highly desirable, which may offer an improved diagnostic and prognostic capability and the development of effective therapeutic strategies.In this study, we detected that not only the expression of miR-93in HCCs significantly increased, but statistical analysis showed that there exists negatively notable correlation between the high expression of miR-93and short overall survival(OS)、short disease free survival(DFS) and high recurrence rate. Thus, miR-93is identified as a prognostic predictor for HCC patients. To identify the biological function of overexpressed miR-93in the progression of HCC, we demonstrated in vitro and in vivo. HCC and normal liver cell line infected with lentivirus mediated miR-93(Lenti-miR-93) showed increased cell growth、cloning formation、mobility and invasion capability. Reversely, both cell lines showed inhibitory effects in vitro. HCC and normal liver cell line infected with Lenti-miR-93injected in nude mice showed a rapid growth and larger volume of tumor compared with those infected with Lenti-anti-miR-93in vivo. Through dual luciferase assay and western blot, EphA4(Ephrin type-A receptor4) was identified as the direct down stream target of miR-93. Knockdown of EphA4phenocopied the effect of miR-93and ectopic expression of EphA4restored the effect of miR-93on proliferation, migration, invasion and adhesion in HCC cells. We further demonstrated that miR-93and EphA4regulated the epithelial-mesenchymal transitioin (EMT) process to affect cell growth and invasion. We determined the expression of E-cadherin,Vimentin, β-catenin and Claudin.Our findings revealed that the expression of miR-93increases in HCCs and may be a survival predictor for HCC following LT (liver transplantation). In addition, we highlight the importance of miR-93in regulating the metastatic properties of HCC by directly targeting EphA4. This may provide new insights into the recurrence mechanism of HCC following LT.