Effect of small interfering RNA transfection on FAK and DLC1 mRNA expression in OVCAR-3

Abstract

RNA interference is an evolutionarily conserved cellular defense mechanism that protects cells from hostile genes and regulates the function of normal genes during growth and development. In this study, we established GFP-siFAK-DLC1 vector and transfect the vector into OVCAR-3 cells. RT-PCR and western blot analyses were performed for FAK, DLC1 mRNA, and protein expression in OVCAR-3 cells. ELISA method was used for caspase-3 and caspase-9 activities. These studies demonstrate that both recombinant pGFP-siFAK-DLC1 vector and pGFP-siCon-DLC1 vector may effectively promote DLC1 mRNA transcription and didn't affect siRNA effect. Recombinant vector (pGFP-siFAK-DLC1) may promote DLC1 gene expression, and effectively silence FAK gene expression. Silencing FAK mRNA expression and DLC1 mRNA expression may markedly enhance caspase-3 and caspase-9 activities in OVCAR-3 cells. These results showed that in ovarian cancer OVCAR-3 cell silencing FAK gene expression or / and increasing DLC-1 gene expression, could improve Caspase-3 and Caspase-9 protease activities. In the expression of DLC-1 and silence FAK expression group (double action group) effect was more significant as compared with the silence FAK gene group or expression of DLC-1 gene alone, difference was significant (p < 0.05).