Staurosporine induces apoptosis in primary hepatocytes by the activation of interleukinlβ converting enzyme (ICE)-like proteases

Carole Couel, Salmaan H. Inayat-Hussain, Kelvin Cain

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Abstract

Interleukin-1β converting enzyme (lCE)-like (e.g. CPP-32, Mch3α) proteases are believed to play a key role in apoptosis and recently we have shown that benzyloxycarbonyl-Val-Ala-Asp-tluoromethylketone (ZVAD.FMK) a selective ICE-like protease inhibitor blocks apoptosis in hepatocyte cultures. In this study we show that staurosporine induces both necrosis and apoptosis in mono-layer cultures of hepatocytes, when cells are cultured for 4 hours before addition of staurosporine. Only the apoptotic effects of staurosporine are blocked by Z-VAD.FMK, indicating that staurosporine induces necrosis and apoptosis by two separate biochemical pathways. Further evidence for this conclusion comes from experiments in which the cells were cultured for 8 hours instead of 4 hours before adding staurosporine. Under these conditions, staurosporine induced apoptosis without necrosis and the apoptotic effect was blocked by Z-VAD.FMK. To further investigate the role of ICE-like proteases in hepatocyte apoptosis we assayed the proteolytic activity of lysates isolated from apoptotic hepatocytes using the model substrate benzoyloxycarbonyl-Asp-Glu-ValAsp-amino-trifluoromethylcoumarin (Z-DEVD.AFC) which is cleaved by CPP-32 like proteases. Lysates from staurosporine treated hepatocytes expressed a marked and time-dependent increase in cleavage aclivity which was totally absent in lysates isolated from hepatocytes which had been pretreated with ZVAD.FMK before the addition of staurosporine. In addlion the increase in cleavage activity proceeded the morphological characteristics of apoptosis. In conclusion, staurosporine induces necrosis and apoptosis by two separate biochemical pathways and in the case of apoptosis involves the activation of ICE (CPP32)-like proteases.

Original languageEnglish
JournalBiochemical Society Transactions
Volume24
Issue number4
Publication statusPublished - 1996
Externally publishedYes

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Staurosporine
Hepatocytes
Peptide Hydrolases
Chemical activation
Apoptosis
Enzymes
Necrosis
aspartylglutamate
Cultured Cells
Caspase 1
Protease Inhibitors

ASJC Scopus subject areas

  • Biochemistry

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Staurosporine induces apoptosis in primary hepatocytes by the activation of interleukinlβ converting enzyme (ICE)-like proteases. / Couel, Carole; Inayat-Hussain, Salmaan H.; Cain, Kelvin.

In: Biochemical Society Transactions, Vol. 24, No. 4, 1996.

Research output: Contribution to journalArticle

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