[Effect of Bax translocation on TNFalpha induced apoptosis of human breast cancer MCF7 cells stably expressed hGFP-Bax].
Established human breast cancer MCF7 cell line stably expressed GFP-Bax, which was used to investigate Bax translocation from cytosol into mitochondria induced by TNFalpha. Bax translocation from cytosol into mitochondria and DNA fragmentation were investigated under fluorescence microscope. The viability of cells treated with TNFalpha was measured by MTT method. Immunofluoresence method was used to investigate Bax translocation to mitochondria and Cyt-c release from mitochondria. The Dose-effect and time-course relationship were also observed and calculated by the percentage of GFP-Bax punctate cells. The TMRE assay was used to investigate the function of mitochondria. The results showed that Bax translocation from cytosol into mitochondria was associated with apoptosis induced by TNFalpha in hGFP-Bax-tagged MCF7 stable cell line.
Hou
Qi
Q
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China. houq@imm.ac.cn
D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D003600:Cytosol / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008928:Mitochondria / Q000378:metabolism*; D021381:Protein Transport; D013995:Time; D014409:Tumor Necrosis Factor-alpha / Q000008:administration & dosage / Q000494:pharmacology*; D051028:bcl-2-Associated X Protein / Q000378:metabolism*
D014409:Tumor Necrosis Factor-alpha; D051028:bcl-2-Associated X Protein; D049452:Green Fluorescent Proteins