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《Journal of Molecular Cell Biology》 2007-01
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CADMIUM INDUCED APOPTOSIS OF HEK293 CELLS AND ITS MITOCHONDRIAL APOPTOSIS PATHWAY

YE Ji Lin MAO Wei Ping WU Ai Lian ZHAO Jie Ming ZHANG Chao ZHANG Na Na JIANG Ping TIAN Ting (College of Life Science,Nanjing Normal University,Nanjing 210097,China)  
HEK293 cell was chose to study the kidney damage of cadmium and to explore the significance of caspase 3,Bcl-2 and AIF(apoptosis inducing factor)in the apoptosis of cells in- duced by cadmium.Inhibition of the cell proliferation was measured by MTT assay.The structure of apoptotic cells was observed by light microscopy and electron microscopy;moreover,apoptotic cells were detected by DNA electrophoresis,flow cytometry and confocal laser microscopy.Further- more,the expressions of Pro-caspase-3,Bcl-2 and the location of AIF in cells(mitochondria,cyto- plasm or nuclei)were tested by western blot and immunofluorescence assay.CdCl_2 exhibited anti- proliferative activity in dosage and time-dependent manner.DNA ladders of HEK293 cells were showed on agarose gel electrophoresis and the fragments of DNA were integral of 180-200 bp.6- 9 hours after 30μmol/L CdCl_2 treatment,DNA ladders were distinct.However,mistiness DNA ladder or smear was found when HEK293 cells were treated with CdCl_2 on higher concentration or treat- ed longer.It suggests that necrosis may happen,and flow cytometry results confirmed it.Morpho- logical examination showed cell shrinkage,chromosomal condensation,karyotheca margination,nu- cleus cracking,vacuoles formed in cytoplasm and the presence of apoptotic bodies.At the same time,mitochondrial membrane potential(MMP)decreased,and the expression of Pro-caspase-3, Bcl-2 were decreased in time-dependent manner.Furthermore,AIF was released from mitochon- dria,and then traveled to nuclei.It suggests that CdCl_2 may induce the apoptosis of HEK293 cells involving mitochondrial disruption including AIF migration and Cyt c release through both caspase- independent and-dependent pathways,and Bcl-2 and Caspase-3 are important factors which partic- ipate in the processes.
【Fund】: 江苏省教育厅自然科学基金资助(2005104TSJB157)。
【CateGory Index】: Q27
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