【摘要】 目的研究SIAH1对乳腺癌细胞失巢凋亡的影响及其相关机制。方法采用脂质体介导法将SIAH1表达质粒pcDNA3-myc-SIAH1、对照质粒pcDNA3-myc、Bim干扰片段BimsiRNA和对照片段control siRNA转染入乳腺癌细胞系MCF-7,利用Western blot法检测SIAH1和Bim的表达,并应用Annexin V-FITC/PI试剂盒及流式细胞仪技术分析各组细胞在悬浮和贴壁生长状态下细胞的凋亡情况。结果与未处理组及转染对照质粒组相比,转染SIAH1表达质粒后,SIAH1(P<0.05)和Bim(P<0.05)表达均明显上调,且失巢凋亡率明显增加,分别为7.36%、8.02%及38.4%。另外,与单独转染SIAH1表达质粒pcDNA3-myc-SIAH1相比,共转染pcDNA3-myc-SIAH1与Bim siRNA后,Bim表达显著降低(P<0.05),同时细胞失巢凋亡率明显降低(P<0.05),分别为36.8%与8.16%。结论过表达SIAH1可通过上调Bim表达来诱导乳腺癌细胞失巢凋亡。更多还原

【Abstract】 Objective To investigate the effect and mechanisms of SIAH1 on the induction of anoikis of breast carcinoma.Methods MCF-7 cells were transfected with pcDNA3-myc-SIAH1,pcDNA3-myc,Bim siRNA or control siRNA using Lipofectamine 2000.The expression of SIAH1 and Bim were detected by Western blot.Annexin V-FITC/PI reagent kit and Flow cytometry assay were used to analyze apoptosis in adherent and nor adherent cells.Results Compared with the control and the cells transfected with the pcDNA3 myc,the level of SIAH1 and Bim were increased in the cells transfected with the pcDNA3-myc-SIAH1.And the anoikis percentage was 7.36%,8.02% and 38.4%,respectively.Further,MCF-7 cells co-transfected pcDNA3-myc-SIAH1 and Bim siRNA showed lower level of Bim(P<0.05) and apoptosis rate(8.16% and 36.8%,respectively) than cells transfected with pcDNA3-myc-SIAH1 only.Conclusion Overexpression of SIAH1 induced anoikis of breast carcinoma by upregulating the level of Bim.更多还原