RNA interference targeting gene of RING finger protein inhibits proliferation and migration of pancreatic cancer cell line PANC-1 / 基础医学与临床

ABSTRACT

Objective To investigate the effect of siRNA-mediated silencing of RNF2 on cell proliferation , migra-tion, cell cycle and apoptosis in human pancreatic cancer PANC-1 cells and its possible mechanism .Methods The siRNA interference was used to down-regulate RNF2 expression.Meanwhile, there were also empty transfection group whose cells were transfected with the control siRNA and mock group without any treatment .The result of transfection was evaluated by fluorescence microscope .The expression of RNF2 mRNA was detected by RT-qPCR. Western blot was applied to detect the expression of RNF 2 and p53.Cell proliferation and migration were analyzed by MTS assay and cell scratch assay , respectively .The transient transfection efficiency , apoptosis rate and cell cy-cle were measured by flow cytometry .Results Compared to the normalized human pancreatic duct epithelial cells , RNF2 expression in pancreatic cancer cells were higher ( P<0.05) .The expression of RNF2 mRNA and protein was decreased in PANC-1 cells by siRNA-RNF2 at 48 h post-transfection.Transfection with siRNA-RNF2 inhibited the proliferation and migration of PANC-1 cells (P<0.05), induced cell apoptosis (P<0.05), increased cell counts in phase G0/G1 and decreased in S and G2/M phase (P<0.05).What's more, after siRNA-RNF2 transfec-tion, the expression of p 53 protein was decreased .Conclusions siRNA-RNF2 can specifically knockdown the ex-pression of RNF2 gene and then inhibit the proliferation and migration of PANC-1 cells.These results indicate RNF2 may be a potential target of gene therapy for pancreatic cancer .