RNA interference targeting EGFR on proliferation of human ovarian carcinoma cell line SKOV_3 / 第三军医大学学报

ABSTRACT

Objective To explore the effect of down-regulating epidermal growth factor receptor (EGFR) expression by RNA interference on the proliferation of ovarian carcinoma SKOV3 cells. Methods The recombinant eukaryotic expression plasmids pGenSil-HK,pGenSil-EGFR1 and pGenSil-EGFR2 were constructed and transfected into SKOV3 cells respectively. The mRNA and protein expressions of EGFR in SKOV3 cells were detected by RT-PCR and Western blotting,respectively. The cell cycle and apoptosis were evaluated by flow cytometry. The proliferation of SKOV3 cells was determined by clone formation assay and MTT assay. Results We successfully constructed the recombinant eukaryotic expression plasmids pGenSil-HK,pGenSil-EGFR1 and pGenSil-EGFR2 and transfected into SKOV3 cells. Three cell clones were screened by G418. Compared with untransfected SKOV3 cells and SKOV3 cells transfected with pGenSil-HK,the expressions of EGFR in SKOV3 cells transfected with pGenSil-EGFR1,pGenSil-EGFR2 were inhibited significantly at both mRNA and protein levels,with an inhibitory rate of 41.87% and 68.07% for EGFR mRNA and of 45.21% and 70.25% for EGFR protein respectively. Compared with untransfected SKOV3 cells and SKOV3 cells transfected with pGenSil-HK,the cell apoptotic rate was significantly increased significantly,the cell cycle was arrested in G1 phase and S phase decreased significantly in pGenSil-EGFR2 SKOV3 cells (P