• Acta Pharmacol. Sin. · May 2002

    Caspases promoted DADAG-induced apoptosis in human leukemia HL-60 cells.

    • Jin-Nan Yang, Chun-Xia Liu, Hua Xu, and Qi-Chao Pan.
    • Department of Pharmacology, Xinxiang Medical College Xinxiang 453003, China. yangjinnan@263.net
    • Acta Pharmacol. Sin. 2002 May 1; 23 (5): 461-6.

    AimTo investigate the roles of caspases in diacetyldianhydrogalactitol (DADAG)-induced apoptosis in human leukemia HL-60 cells.MethodsInhibition 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.ResultsDADAG 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.ConclusionDADAG induced apoptosis in HL-60 cells by activating caspases. Caspases promoted apoptosis through the cleavage of substrates of PARP, lamin B, and DFF45.

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