Effect of retrovirally mediated RNA in-terference targeting human Bmi-1 gene on MCF-7 cell line
AIM: To investigate the inhibition effects of Bmi-1 (B-cell-specific moloney routine leukemia virus insertion site 1) with RNA interference (RNAi) on the apoptosis and proliferation of breast carcinoma MCF-7 cells. METHODS: Recombinant retroviral Bmi-1 gene short hairpin RNA(shRNA) expression vec-tor pSuper-retro/Bmi-1 si was transiently transfected into breast carcinoma MCF-7 cells and pSuper-retro/GFP si vector was used as negative control. Bmi-1 mRNA and protein of the transfected cells were examined with reverse transcriptianal polymerase chain reaction(RT-PCR) and Western blot assay, respectively. The proliferations of MCF-7 cells were examined by MTr and the cell cycle status and apoptosis of MCF-7 cells were detected by flow cytometry (FCM). RESULTS : pSuper-retro/Bmi-1 si retroviral vector resulted in effective inhibition of Bmi-1 mRNA and protein. Compared with that in uninfected control and negative control, no significant increase was observed in the basal levels of apoptosis in MCF-7 cells infected with pSuper-retro/Bmi-1 si. G1/S phase transformation and the growth of MCF-7 cells were significantly inhibited by Bmi-1. CONCLUSION: Retroviral shRNA expres-sion vector pSuper-retro/Bmi-1 si significantly inhibits the expres-sion of Bmi-1 and exerts effective inhibition effect on the viability of MCF-7 cells. The results of this study provide evidence for gene therapy for human breast cancers.
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万方数据
维普
