Activation of actin-cleavable interleukin 1beta-converting enzyme (ICE) family protease CPP-32 during chemotherapeutic agent-induced apoptosis in ovarian carcinoma cells.
We have previously reported that actin cleavage activity (ACA) by interleukin 1beta-converting enzyme (ICE) family protease was elevated during anticancer drug-induced apoptosis in human leukemia U937 cells. In this study, the involvement of ACA in the drug-induced apoptosis in solid tumor cells was investigated. Human ovarian carcinoma OVCAR-3 cells undergo apoptotic cell death when cells are treated with chemotherapeutic agents such as cisplatin and etoposide. The induction of the actin cleavage activity accompanied the development of apoptosis. ICE/ced-3 family protease inhibitors such as Z-VAD-CH2DCB and Z-EVD-CH2DCB at 100 microg/ml prevented both the emergence of ACA and the morphological change, characteristics of apoptosis, in cisplatin-treated OVCAR-3 cells. The ACA in apoptotic OVCAR-3 cell lysate was greatly adsorbed by antibody against CPP-32, an ICE family protease. Furthermore, the immunoprecipitated CPP-32 from OVCAR-3 lysate could cleave actin to generate a 15-kDa fragment, as did the apoptotic OVCAR-3 cell lysate, indicating that CPP-32 is a major protease responsible for the ACA. The activation of CPP-32 in the drug-treated cell lysate was verified with Western blot analysis. Our present results indicate that CPP-32, an actin cleavage ICE/ced-3 family protease, could be a common mediator involved in the process of chemotherapy-induced apoptosis of cancer cells.
Laboratory of Biomedical Research, Institute of Molecular and Cellular Biosciences, The University of Tokyo, Bunkyo-ku, Japan.
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Z
Chen
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Naito
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Mashima
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Tsuruo
D000199:Actins / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000235:genetics; D020170:Caspase 1; D002945:Cisplatin / Q000494:pharmacology; D003546:Cysteine Endopeptidases; D053938:DNA Fragmentation; D004273:DNA, Neoplasm / Q000187:drug effects*; D005047:Etoposide / Q000494:pharmacology; D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000188:drug therapy / Q000201:enzymology* / Q000473:pathology / Q000648:ultrastructure; D011480:Protease Inhibitors / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured
D000199:Actins; D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; D011480:Protease Inhibitors; D005047:Etoposide; D003546:Cysteine Endopeptidases; D020170:Caspase 1; D002945:Cisplatin