MiR-10a Is Involved In Migration And Invasion Of Hepatoma Cells

Objective:The processes of migration and invasion are cascade of linked sequential steps involving multiple regulated genes, the crypticity and predictability are the most insidious and life-threatening aspects of cancer. Our understanding of the processes of migration and invasion has improved, and the discovery of microRNAs(miRNAs) explores a new way to find the answer. miRNAs are the new family of small RNA molecules, which are proved to negatively regulate certain genes containing complementary sequences in3’untranslated regions. In several cases, miRNAs have shown to affect target genes involved in cell proliferation, apoptosis, differen-tiation, metabolism. The objective of this paper is to find the miRNAs contributing to the progression of migration and invasion, and to reveal the mechanism through confirmation of target genes.Methods:Our lab had proved that miR-lOa was upregulated in low metastasis colon cancer, and was a key regulator of metastasis in colon cancer. MMP14was identified experi-mentally as a target gene of miR-10a. So we did several experiments to detect the role of miR-10a in hepatic cancer. By wound-healing assay and transwell, we measured the migration and invasion abilities of hepatic cancer cell after miR-10a was blocked or upexpressed. Target genes of miR-10a were predicted by Bioinformatics. Then we chose a gene(EPHA4) which was involved in the process of metastasis. By EGFP reporter assay the predicted gene was verified experimentally, by Real-time PCR and Western blot the mRNA and protein levels of target gene were tested. The cell was transfected with plasmid pSilenser-EPHA4then tested the invasion ability.Results:The migration and invasion abilities of hepatic cancer cell were elevated dramatically after miR-10a was high-expressed. EPHA4was the putative target gene of miR-10a predicted by computational target prediction algorithm. EGFP reporter carrying the3’untranslated regions of EPHA4can be inhibited by miR-10a. The results of Real-time PCR and Western blot shown that miR-lOa could impact the expression of EPHA4on mRNA and protein levels.Conclusion:We confirmed that miR-lOa is a key regulator of migration and invasion in hepatic cancer. EPHA4plays a role in migration and invasion of hepatic cancer as the one of its target gene.