南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (09): 1285-.

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人凋亡诱导因子C-末端截短体AIFΔ1-480 的表达对MCF-7细胞的影响

王丽娟，肖华卫，白娥，周霞，李萌，金桂花，朱青   

  • 出版日期:2013-09-20 发布日期:2013-09-20

Effect of the C-terminal truncated human apoptosis-inducing factor Δ1-480 on biological behaviors of MCF-7 cells

  • Online:2013-09-20 Published:2013-09-20

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摘要： 目的观察人亡诱导因子（AIF）C-末端端截短体AIFΔ1-480重组基因表达对MCF-7细胞生物学行为的影响。方法利用
脂质体将重组质粒PcDNA3.0-FDT- AIFΔ1-480瞬时转染乳腺癌MCF-7细胞，通过Western blot、流式细胞术、MTT、集落形成试
验、线粒体膜电位测定等方法，检测目的基因在细胞中的表达及其对转染细胞凋亡相关分子cytC 表达的变化及细胞增殖、凋亡
等的影响。结果重组质粒PcDNA3.0-FDT-AIFΔ1-480 转染肿瘤细胞后AIF蛋白表达明显增加；重组质粒能明显抑制乳腺癌
MCF-7 细胞增殖；上调细胞色素C的表达；减低线粒体膜电位，正常细胞、空载体组和重组质粒的线粒体膜电位（Δψ）分别为
4.69±1.07、3.86±1.52和1.51±0.79 mV，重组质粒组与对照组相比，线粒体膜电位明显下降（P<0.05）；并诱导细胞凋亡，正常细胞
组、脂质体组、PcDNA3.0组、重组质粒组细胞凋亡率分别为：（9.2±5.1）%、（15.4±4.5）%、（20.5±3.9）%、（32.2±6.0）%，重组质粒组与
对照组相比，细胞凋亡明显增加（P<0.05）。结论人AIF C-末端端截短体可通过诱导cytC从线粒体中释放促进MCF-7细胞凋亡。

Abstract: Objective To observe the expression of the C-terminal truncated human apoptosis-inducing factor (AIF) and its
biological effect on MCF-7 cells. Methods PcDNA3.0-FDT-AIFΔ1-480 was transfected into human breast carcinoma MCF-7
cells with lipofectamins. The expression of the truncated AIF gene was detected by Western blotting, and its effects on the
biological behaviors of MCF-7 cells and on the expression of cytochrome c (cytC) were evaluated using flow cytometry, MTT
assay, colony-forming assay, and mitochondrial membrane potential measurement. Results PcDNA3.0-FDT-AIFΔ1-480
enhanced AIF expression in MCF-7 cells, obviously inhibited the cell proliferation, and significantly reduced the mitochondrial
membrane potentials (P<0.05). Transfection of the cells with PcDNA3.0-FDT-AIFΔ1-480 promoted the expression of cytC and
resulted in significantly increased apoptosis of MCF-7 cells (P<0.05). Conclusion The expression of C-terminal truncated
human AIF gene can induce apoptosis of human MCF-7 cells by promoting cytC release from mitochondria.