Specific siRNA targeting EGFR enhances ovarian cancer cell line SKOV-3 apoptosis

Abstract

Objective To observe the influence of specific short hairpin siRNA targeting EGFR gene on apoptosis of human ovarian cancer Skov-3 cells in vitro. Methods A plasmid of a short hairpin siRNA targeting EGFR was constructed, and it was transfeeted 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 small hairpin RNA expression vector. The expression of EGFR mRNA and protein were examined by RT-PCR and immunocytochemistry, Flow cytometry (FCM) was adopted to analyze quantitatively apoptotic cells in each group. Results After transfection of pshRNA-EGFR, mRNA and protein levels of EGFR gene in Skov-3 cells were obviously reduced. Flow cytometry analysis revealed that apoptosis could be induced in Skov-3 cells line transfected with pshRNA-EGFR in a time-dependent manner, no obvious apoptosis were detected in control group and non-specific group. Conclusion The plasmid expressive vector target at EGFR in our study is capable of suppressing EGFR expression of human ovarian cancer Skov-3 cells and inducing apoptosis, which provide a new way for the gene therapy of human ovarian cancer. Key words: Genes,erbB-1; Ovarian neoplasms; RNA interference; Apoptosis