[Apoptosis induced by diacetyldianhydrogalactitol and its mechanism in HL-60 leukemia cells].
OBJECTIVE
To investigate the apoptosis induced by diacetyldianhydrogalactitol (DADAG) and its mechanism in human HL-60 leukemia cells.


METHODS
Inhibition of proliferation was measured by MTT assay. DADAG-induced apoptosis in HL-60 cells was observed by electron microscopy, flow cytometry and DNA fragmentation assay. The levels of Bcl-2 family proteins were detected by Western blotting. Caspase-3 activity was determined by ApoAlert CPP32 colorimetric assay kit.


RESULTS
DADAG exhibited potent antiproliferative activity and induced apoptosis in HL-60 cells. After treatment with DADAG 8 micrograms.mL-1 for various times, the Bcl-XL protein level decreased in a time-dependent manner, while the Bad protein level was upregulated. The caspase-3 activity increased markedly after treatment with DADAG for 24 h. The apoptotic signals were suppressed by z-VAD.fmk (a general inhibitor of caspases), whereas z-DEVD.fmk, a selective inhibitor of caspase-3, only induced partial reversion of the apoptotic effects.


CONCLUSIONS
DADAG-induced apoptosis in HL-60 cells required caspase-3 activation and caspase-3 activation was related with Bcl-2 family members.
Yang
Jin-nan
JN
Department of Pharmacology, Xinxiang Medical College, Xinxiang 453003, China. yangjinnan@263.net
Liu
Ju-yuan
JY
Xu
Hua
H
Liu
Xiao-li
XL
Qin
Yu
Y
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D003961:Dianhydrogalactitol / Q000031:analogs & derivatives* / Q000494:pharmacology*; D018922:HL-60 Cells; D006801:Humans; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051020:bcl-X Protein
D000970:Antineoplastic Agents; C494811:BCL2L1 protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D051020:bcl-X Protein; D003961:Dianhydrogalactitol; C027991:dianhydro-3,4-diacetylgalactitol; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases