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目的:探讨靶向SATB1(special AT-rich sequence-binding protein-1)基因的短发夹RNA(short hairpin RNA,shRNA)对肺癌A549细胞凋亡的影响及其可能的机制。方法:构建靶向SATB1基因的shRNA重组质粒SATB1-shRNA,采用脂质体法将其转染至A549细胞;分别采用RT-PCR和蛋白质印迹法检测A549细胞中SATB1、Bcl-2、BaxmRNA和SATB1、Bcl-2、Bax、caspase3蛋白的表达水平;FCM检测A549细胞的凋亡率。结果:成功构建了SATB1-shRNA重组质粒;SATB1-shRNA转染A549细胞后,SATB1、Bcl-2 mRNA及其蛋白表达下调,Bax mRNA和Bax、caspase3蛋白表达上调(P<0.05)。SATB1-shRNA转染组细胞凋亡率[(14.18±1.59)%]较对照组[(1.84±0.57)%]明显增加(P<0.01)。结论:SATB1-shRNA可显著下调肺癌A549细胞中SATB1基因的表达水平并诱导细胞凋亡,其机制可能与下调Bcl-2基因表达所引起的级联效应有关。
Objective: To investigate the effect of short hairpin RNA (shRNA) targeted to the special AT-rich sequence-binding protein-1 gene on the apoptosis of lung cancer A549 cells and its possible mechanism. METHODS: SATB1 shRNA targeting SATB1 gene was constructed and transfected into A549 cells by lipofectamine 2000. SATB1, Bcl-2, Bax mRNA and SATB1 were detected by RT-PCR and Western blot respectively , Bcl-2, Bax, caspase3 protein expression; FCM A549 cell apoptosis rate. Results: The SATB1-shRNA recombinant plasmid was successfully constructed. The expression of SATB1 and Bcl-2 mRNA and protein was down-regulated while the expression of Bax and Bax and caspase3 were up-regulated in A549 cells transfected with SATB1-shRNA (P <0.05). The apoptosis rate of SATB1-shRNA transfected group [(14.18 ± 1.59)%] was significantly higher than that of control group [(1.84 ± 0.57)%] (P <0.01). Conclusion: SATB1-shRNA can significantly down-regulate the expression of SATB1 gene and induce apoptosis in A549 cells. The mechanism may be related to the cascade effect induced by the down-regulation of Bcl-2 gene expression.