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Pro-apoptotic activity of truncated human apoptosis-inducing factor on HeLa cells
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 7
- DOI:
- 10.3321/j.issn:1000-2790.2007.07.001
- Key Word:
- 凋亡诱导因子;HeLa细胞;细胞凋亡
Abstract： 目的:观察人截短型凋亡诱导因子(AIF△1-400)对HeLa细胞的促凋亡作用.方法:在AIF△1-120基因克隆成功的基础上进一步改造,构建截去AIF基因线粒体定位信号,FAD结合结构域和NADH结合结构域(1-400位氨基酸)编码序列的AIF△1-400基因,将其克隆入pcDNA3和pIRES2-EGFP真核表达载体,用脂质体法转染HeLa细胞,通过荧光显微镜观察,MTT检测等方法检测该基因在转染细胞中的表达及其对肿瘤细胞的促凋亡活性.结果:经酶切鉴定与测序证实,带有AIF△1-400的真核表达载体构建成功.瞬时转染后出现细胞形态变化,随转染时间的延长转染细胞数减少,荧光变弱.经间接免疫荧光检测可观察到截短型AIF蛋白位于细胞质中,部分已位于细胞核内.MTT法检测发现AIF△1-400转染后对细胞生长有明显的抑制作用.结论:AIF△1-400仍然具有诱导HeLa细胞凋亡的作用.
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