RNA干扰抑制MTA 1的表达对乳腺癌细胞MDA-MB-231增殖的影响

目的:探讨通过RNA干扰技术抑制MTA1的表达及对乳腺癌细胞MDA-MB-231增殖和细胞周期的影响。方法:建立3种稳定表达靶向MTA1的shRNA的MDA-MB-231细胞系。通过反转录-聚合酶链反应(reverse transeription-polymerase chain reaction,RT-PCR)和免疫组化法验证基因表达的抑制效果;采用细胞计数法及噻唑蓝(thiazolylblue,MTT)法检测细胞的增殖能力;流式细胞仪分析细胞周期。结果:MTA1基因被抑制后,其mRNA表达明显下降,RT-PCR结果显示,稳定转染3种重组质粒后,MTA1mRNA水平较对照组分别下调了70.07%、82.40%和63.44%;同时,MDA-MB-231细胞的生长明显受抑制,与对照组细胞相比,细胞计数示细胞数目明显减低;MTT分析示细胞增殖下降,流式细胞分析示细胞周期阻滞在G0/G1期。结论:在MDA-MB-231细胞中,抑制MTA1的表达可有效抑制细胞生长并导致细胞周期的阻滞;利用RNA干扰技术抑制MTA1的表达可能会成为一种有效的乳腺癌基因治疗手段。

Effects of suppression of MTA1 expression by RNA interference on proliferation of human breast carcinoma cell line MDA-MB-231

Objective:To explore the effects of suppressing MTA1 expression using RNA interference technique on the proliferation and cell cycle of human breast carcinoma cell line MDA-MB-231.Methods:We established three MDA-MB-231 cell lines which had stable expression of MTA1-targeted shRNA.The suppression of MTA1 expression was verified by RT-PCR and immunohistochemistry.Cell proliferation capacity was evaluated by cell counting and MTT assay.The cell cycle was analyzed by flow cytometry.Results:Expression of MTA1 mRNA was significantly down-regulated after silencing MTA 1 gene by RNA interference.RT-PCR results showed that the expression of MTA1 mRNA was down-regulated by 70.07%,82.40%,63.44% in MDA-MB-231 cells after stable transfection of three types of plasmids.The growth of MDA-MB-231 cells was inhibited significantly.Compared with the control group,the cell number significantly decreased.MTT assay and flow cytometry showed that cell proliferation capacity decreased and the cells were arrested at G0/G1 phase after silencing MTA 1 gene.Conclusion:Suppression of the expression of MTA1 in MDA-MB-231 cells effectively inhibited cell proliferation and induced cell cycle arrest.Silencing MTA 1 gene by RNA interference may be a useful approach of gene therapy for human breast carcinoma.