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Inhibitors of cysteine cathepsin and calpain do not prevent ultraviolet-B-induced apoptosis in human keratinocytes and HeLa cells

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Abstract

Caspases, members of the cysteine protease family, execute UVB-induced apoptosis in several cell lines and keratinocytes. Several researchers investigating UVB-induced apoptosis have demonstrated a dose-dependent protective effect of the synthetic peptide caspase inhibitor zVAD-fmk. However, zVAD-fmk displays a dose-dependent protective effect against UVB-induced apoptosis, even at doses higher than those required to block all known proapoptotic caspases. In addition, it is known that zVAD-fmk also inhibits other cysteine proteases including cathepsins and calpains, and these proteases have recently been demonstrated to play a role in the execution of programmed cell death induced by other stimuli, e.g. TNF-α. The purpose of the present study was therefore to investigate whether inhibitors of cysteine cathepsins and calpains could prevent UVB-induced apoptosis in HeLa cells and keratinocytes. This was done by investigating the effect of the irreversible cysteine protease inhibitor zFA-fmk, the cathepsin B inhibitor CA-074-Me and the calpain inhibitor ALLN on the viability of UVB-irradiated human keratinocytes and HeLa cells. At concentrations of 10 μM and above zVAD-fmk conferred partial dose-dependent protection against UVB-induced apoptosis in HeLa cells and keratinocytes. Moreover, caspase-3 activity was completely blocked at zVAD-fmk concentrations of 1 μM in HeLa cells. This indicates that caspase-independent mechanisms could be involved in UVB-induced apoptosis. However, the protease inhibitors zFA-fmk, CA-074-Me and ALLN all failed to prevent UVB-induced apoptosis in HeLa cells and keratinocytes. In conclusion, the protective effect of zVAD-fmk at high concentrations indicates that other proteases than caspases are active in the execution of UVB-induced apoptosis but further studies are needed to identify these proteases.

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Acknowledgements

We are grateful to Birgit Poulsen for excellent technical assistance. The study was supported by the Copenhagen Council, Aage Bang’s Foundation, the Danish Hospital Foundation for Medical Research. Region of Copenhagen, The Faroe Islands and Greenland, and the Danish Cancer Society.

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Authors and Affiliations

  1. Department of Dermatology, Gentofte Hospital, Copenhagen, Denmark

    Bo Bang, Ole Baadsgaard & Lone Skov

  2. Apoptosis Laboratory, Danish Cancer Society, Copenhagen, Denmark

    Marja Jäättelä

  3. Department of Dermatology D40, Bispebjerg Hospital, University of Copenhagen, Bispebjerg Bakke 23, DK-2200, Copenhagen NV, Denmark

    Bo Bang

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  1. Bo Bang
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Correspondence to Bo Bang.

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Bang, B., Baadsgaard, O., Skov, L. et al. Inhibitors of cysteine cathepsin and calpain do not prevent ultraviolet-B-induced apoptosis in human keratinocytes and HeLa cells. Arch Dermatol Res 296, 67–73 (2004). https://doi.org/10.1007/s00403-004-0473-4

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  • DOI: https://doi.org/10.1007/s00403-004-0473-4

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