Proª²apoptotic activity of truncated human apoptosisª²inducing factor on HeLa cells

    ZHANG Wei, ZHANG Yong, MENG Yanª²Ling, WEN Weiª²Hong, XUE Qian, REN Junª²Lin, YANG Anª²Gang

    Department of Immunology, School of Basic Medicine, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To observe the expression of truncated human apoptosisª²inducing factor (AIF) gene and its effect on HeLa cells. METHODS: After AIF¡÷1-120 gene was successfully cloned, the shorter truncated human AIF gene (AIF¡÷1-400) was constructed by deleting the Nª²terminal mitochondrial localization sequence(MLS), the coding sequence of flavine adenine dinucleotide (FAD) binding domain and nicotinamide adenine dinucleotide (NADH) binding domain of AIF protein.The truncated human AIF gene (AIF¡÷1-400) was inserted into the pIRES2ª²EGFP and pcDNA3 eukaryotic expression vectors, and the coª²expression vectors were then transfected into HeLa cells with LipofectAMINE. The expression of the truncated AIF gene and its effect on HeLa cells were detected by fluorescence microscope analysis and MTT test. RESULTS: The eukaryotic expression vector containing the truncated human AIF (AIF¡÷1-400) gene was successfully constructed. The AIF protein could be detected in the transfected HeLa cells. After transfection, typical cell apoptotic changes were observed under an electron microscope. MTT test showed that the proliferation of HeLa cells was significantly inhibited. CONCLUSION: The expression of the truncated human AIF(AIF¡÷1-400)gene can induce the apoptosis of the transfected human HeLa cells. ......