Inhibition of A549 cell proliferation by RNA interfered gene her2/neu

Objective:The effect of HER2/neu expression on the proliferation of A549 cell is not clear. We inhibited the HER2/neu expression with RNA interference (RNAi) technique,and investigated the proliferation of A549 cell in this study. Methods:Firstly a recombinant RNAi expression plasmid targeting to her2/neu gene (named psilence 3.1-HER2) was constructed. After RNAi plasmids being transfected to A549 cells with Lipofectamine,the positive clones were screened with G418. The expression changes of HER2/neu mRNA and protein were evaluated with RT-PCR and Western Blot respectively; Also FCM analysis,MTT method were applied to measure cell cycle,cell growth of A549 cell respectively after the transfection of psilence 3.1-HER2. Results:The psilence 3.1-HER2 was successfully constructed and can specially inhibit HER2/neu mRNA and protein expression after transfection. FCM results showed that cells transfected with psilence 3.1-HER2 increased accumulation of cells in G0/G1 phase with a decrease in the percentage of cells in S-phase,and the growth curve of cells exhibited slower proliferation. Conclusion:The psilence 3.1-HER2 can knockdown her2/neu gene specially and efficiently,transfection of psilence 3.1-HER2 contributes to the depression of A549 cell proliferation. The data described here provided a proof of the concept that vector-based RNAi targeting to her2/neu gene could be used therapeutically for lung cancer.