Caspases promoted DADAG-induced apoptosis in human leukemia HL-60 cells.
OBJECTIVE
To investigate the roles of caspases in diacetyldianhydrogalactitol (DADAG)-induced apoptosis in human leukemia HL-60 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. Caspase-3 activity was determined by ApoAlert CPP32 colorimetric assay kit. The cleavage of substrates of caspases was detected by Western blot.


RESULTS
DADAG exhibited potent antiproliferative activity and induced apoptosis in HL-60 cells. After treatment with DADAG 8 mg/L for 24 h, caspase-3 activity increased markedly and the cleavage of poly-(ADP-ribose) polymerase (PARP), lamin B, and DFF45 appeared. All of the apoptotic signals were suppressed by z-VAD fmk (a general inhibitor of caspase activities), whereas z-DEVD fmk, a selective inhibitor of caspase-3 activity, only induced partial reversion of the apoptotic effects.


CONCLUSIONS
DADAG induced apoptosis in HL-60 cells by activating caspases. Caspases promoted apoptosis through the cleavage of substrates of PARP, lamin B, and DFF45.
Yang
Jin-Nan
JN
Department of Pharmacology, Xinxiang Medical College  Xinxiang 453003, China. yangjinnan@263.net
Liu
Chun-Xia
CX
Xu
Hua
H
Pan
Qi-Chao
QC
D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism* / Q000502:physiology; D002455:Cell Division / Q000187:drug effects; D003961:Dianhydrogalactitol / Q000031:analogs & derivatives* / Q000494:pharmacology*; D018922:HL-60 Cells; D006801:Humans; D007938:Leukemia / Q000473:pathology
D003961:Dianhydrogalactitol; C027991:dianhydro-3,4-diacetylgalactitol; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases