The Experimental Study Of Invasion And Migration By Down-regulating The Expression Of UBC9 In Hepatocellular Carcinoma Cell Line

Purpose:To investigate the expression of UBC9 in hepatocellular carcinoma cells and research the effects of reducing UBC9 expression by means of short hairpin RNA on the invasion and metastasis in human live cancer and analyze the underlying mechanisms.Methods:Hepatocellular carcinoma cells were separate into four grups: normal hepatoma cells,liposome-transfected cells,non-specific shRNA-transfected cells,shRNA-targeting UBC9-transfected cells.Hepatoma cells were instantaneously transfected using lipofectamine method.Downregulation of UBC9 gene expression in MHCC97 h cells by siRNA.UBC9-shRNA interference effects was detected by Q-PCR and western-blot.Wound healing assay and transwell assay were performed to detect the effects of ubc9 on the invasion and metastasis of hepatocellular carcinoma cells.The expression of ?-catenin,MMP-2 and MMP9 proteins were detected by Western blot assay after transfected cells 48 hours.Results:The mRNA expression and protein expression of UBC9 in the UBC9-shRNA group was significantly lower compared with the control groups.Wound healing assay showed the cell migration ability of UBC9-shRNA group was similarly lower than the other control groups.Transwell assay revealed the number of invaded cells in UBC9-shRNA group were markedly reduced compared with the control groups.Western blot assay displayed the expression of ?-catenin,MMP-2,MMP9 in UBC9-shRNA group reduced obviously compared with the control groups when UBC9 gene expression was down regulated.Conclusions:UBC9 gene plays an important role in the invasion and metastasis of hepatocellular carcinoma.Ubc9 is able to promote the migration and invasion of human MHCC97 H cell line and may be associated with Wnt signal pathway and the downstream genes include MMP-2,MMP-9.