Effect of truncated human apoptosis-inducing factor expression on apoptosis of HeLa cells

AIM: To observe the expression of the truncated human apoptosis-inducing factor (AIF) gene and its apoptosis-inducing effect on HeLa cells.METHODS: After the truncated human apoptosis-inducing factor (AIF△1-120) gene was successfully cloned,the shorter truncated human AIF gene (AIF△1-480) was cloned,which was constructed by deleting the N-terminal mitochondrial location sequence (MLS) and the coding sequence of flavin adenine dinucleotide (FAD) binding domain and nicotinamide adenine dinucleotide (NADH) binding domain of AIF protein.The truncated human AIF gene(AIF△1-480) was inserted into the EGFP co-expression vector pIRES2-EGFP,and the pIRES2-EGFP-AIF△1-480 was then transfected into HeLa cells with LipofectAMINE.The expression of the truncated AIF gene and its effect on HeLa cell were detected by fluorescence microscope and electron microscope analysis.RESULTS: The eukaryotic expression vector pIRES2-EGFP containing the truncated human AIF(AIF△1-480)gene was successfully constructed.The AIF protein could be detected in the transfected HeLa cells.After transfection,typical apoptotic changes of the transfected HeLa cells and a mass of cell death were observed under electron microscope.CONCLUSION: The expression of the truncated human AIF(AIF△1-480)gene can induce apoptosis of the transfected human HeLa cells.