Effect of EGFR Gene Silencing on Cell Cycle and Apoptosis of Ovarian Cancer Cell Line Skov-3

Objective To explore the effect of RNA interference (RNAi)-mediated inhibition on epidermal growth factor receptor (EGFR) gene expression in ovarian cancer cells. Methods The DNA sequence of EGFR gene was synthesized in vitro and cloned into pSilencer 2.1-U6neo plasmid vector to construct the shRNA expression vector. After the constructed vector was transfected into human ovarian cancer Skov-3 cells using lipofectamine 2000, semi-quantitative RT-PCR and immunocytochemistry assay were conducted to compare the changes in the expression levels of EGFR between before and after transfection. Moreover, cells were stained with Annexin V/PI and PI and then siRNA-induced apoptosis and cell cycle analyzed. Results SiRNA eukaryotic expression vector pSilencer-EGFR was successfully constructed; the siRNA targeted against EGFR could effectively inhibit EGFR gene expression in Skov-3 cells; the results of flow cytometric analysis indicated that the apoptosis rate of Skov-3 cells increased significantly and an obvious cell cycle arrest at G0/G1 phase was induced. Conclusion RNAi-mediated silencing of EGFR gene expression can induce cell apoptosis and cell cycle arrest at G0/G1 phase. Thus, RNAi can be used as an effective tool to explore the mechanism underlying the development and progression of ovarian cancer as well as conduct gene therapy of this disease.