Gambogic acid induces death inducer-obliterator 1-mediated apoptosis in Jurkat T cells.
OBJECTIVE
To explore the anticancer effects and the molecular mechanisms of gambogic acid (GA) on Jurkat cells.


METHODS
Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Annexin V-fluorescein-isothiocyanate/propidium iodide, DNA defragmentation, and comet assay were used to detect apoptosis. Western blotting was used to study the expression of death inducer-obliterator-1 (DIO-1), Bcl-2, NF-kappaB, and pro-caspase 3, as well as 2 activated subunits: p17 and p20. The subcellular localization of DIO-1 was examined by immunofluorescence and Hoechst33258 staining.


RESULTS
GA inhibited the proliferation of Jurkat cells with 50% inhibitory concentration values of 1.51+/-0.09 (24 h), 0.98+/-0.13 (48 h), and 0.67+/-0.12 micromol/L (72 h). GA was able to induce apoptosis of Jurkat cells. Treated by GA, the expression of DIO-1 was upregulated, and that of Bcl-2 and NF-kappaB was downregulated, leading to the activation of pro-caspase 3. GA induced the translocation of DIO-1 to the nucleus.


CONCLUSIONS
GA suppressed the proliferation of Jurkat cells by apoptosis induction. DIO-1 triggered early-stage cell death in GA-treated Jurkat cells.
Wang
Yong
Y
Institute of Hematology, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. yanchen@public.wh.hb.cn.
Chen
Yan
Y
Chen
Zi
Z
Wu
Qing
Q
Ke
Wen-juan
WJ
Wu
Qiu-ling
QL
D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D006690:Bisbenzimidazole / Q000378:metabolism; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D005562:Formazans / Q000378:metabolism; D006801:Humans; D020128:Inhibitory Concentration 50; D019169:Jurkat Cells; D015394:Molecular Structure; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D011419:Propidium / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects*; D013778:Tetrazolium Salts / Q000378:metabolism; D044004:Xanthones / Q000737:chemistry / Q000494:pharmacology*
D017304:Annexin A5; C518584:DIDO1 protein, human; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D005562:Formazans; D016328:NF-kappa B; D019253:Proto-Oncogene Proteins c-bcl-2; D013778:Tetrazolium Salts; D044004:Xanthones; C079782:MTT formazan; D011419:Propidium; C052659:gambogic acid; D053148:Caspase 3; D016650:Fluorescein-5-isothiocyanate; D006690:Bisbenzimidazole