SIAH1在乳腺癌中上调Bim的表达诱导乳腺癌细胞凋亡研究

SIAH1 induces apoptosis of breast cancer cells by up-regulating the level of Bim

目的探讨SIAH1在乳腺癌中的表达及对乳腺癌细胞凋亡的影响。方法采用逆转录PCR和免疫印迹试验法检测30例乳腺组织与乳腺癌细胞中SIAH1的表达情况。采用脂质体介导法将SIAH1表达质粒pcDNA3-myc-SIAH1、对照质粒pcDNA3-myc、SIAH1干扰片段SIAH1 siRNA1、对照片段control siRNA1、Bim siRNA1及对照片段control siRNA2转染入乳腺癌细胞系MDA-MB-231后,利用逆转录PCR和免疫印迹试验法检测SIAH1基因在m RNA和蛋白水平的表达情况及对Bim表达的影响,并应用流式细胞仪技术分析检测SIAH1对MDA-MB-231细胞凋亡的影响。结果 SIAH1在乳腺癌组织和细胞中低表达。与乳腺癌细胞系MDAMB-231转染对照质粒组相比,转染pc DNA3-myc-SIAH1后,Bim表达上调,细胞凋亡率增加;干扰SIAH1表达后,Bim表达下调。与乳腺癌细胞系MDA-MB-231转染pcDNA3-myc-SIAH1相比,共转染pcDNA3-myc-SIAH1和Bim siRNA1后,凋亡率明显减少。结论过表达SIAH1可通过上调Bim表达来诱导乳腺癌细胞凋亡。

Objective To investigate the expression of SIAH1 in breast carcinoma and its effect on the induction of breast carcinoma apoptosis. Methods The expression of SIAH1 mRNA and protein were examined in breast cancer tissues and cell lines by RT-PCR and western blot. MDA-MB-231 cells were transfected with pc DNA3-myc-SIAH1,pcDNA3-myc, SIAH1 siRNA1, control si RNA1, Bim siRNA1 or control siRNA2 by using Lipofectamine 2000. The expression of SIAH1 and Bim were detected by RT-PCR and Western blot. SIAH1-inducing apoptosis of breast carcinoma was detected by Flow cytometry assay. Results SIAH1 mRNA and protein expression were significantly lower in breast cancer tissues and cell lines. Compared with the control and the cells transfected with the pc DNA3-myc, the level of SIAH1 and Bim were increased in the cells transfected with the pcDNA3-myc-SIAH1. And the apoptosis percentage was also increased. On the contrary, the level of SIAH1 and Bim were decreased in the cells transfected with the SIAH1 siRNA1. Conclusions Overexpression of SIAH1 can induce apoptosis of breast carcinoma by up-regulating the level of Bim.