Influence of siRNA on the ovarian carcinoma cell cycle and EGFR expression

Objective To establish the small interference RNA(siRNA) plasmid expression vector of coding human epidermal growth factor receptor (EGFR) gene in vitro, and investigate its specific inhibition on EGFR expression in human ovarian cancer Skov-3 cells and the effect of siRNA on cell apoptosis and cell cycle. Methods The plasmid of siRNA targeting EGFR was constructed, and transfected into Skov-3 cell line by lipofectamine 2000. Human ovarian carcinoma cells of the line Skov-3 were cultured and divided into 3 groups: control group; non-specific group, transfected with non-specific plasmid vector; and specific group, transfected with specific siRNA expression vector. The expression of EGFR mRNA and protein were examined by RT-PCR and immuno-fluorescent method. Flow cytometry (FCM) was adopted to analyze apoptotic cells and cell cycle in each group. Results Plasmid expressive vectors aimed directly at EGFR were successfully established. psiRNA-EGFR plasmid knocked down EGFR expression in Skov -3 cells obviously, arrested the cell cycle in G1 phase, and cell apoptosis was promoted. Conclusion The recombinant plasmid psiRNA-EGFR can efficiently and specifically inhibit the expression of EGFR gene and proliferation of Skov-3 cells, the inhibiting mechanism of which may be associated with redistribution of the cell cycle, knocking down the ratio of S phase cell and enhancing apoptosis.