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Introduction of an N-terminal peptide of S100C/A11 into human cells induces apoptotic cell death

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Abstract

S100 proteins belong to the EF-hand Ca2+-binding protein family and are involved in the regulation of a variety of cellular processes. Individual S100 proteins are expressed in cell- and tissue-specific manners, and functional deterioration of S100 proteins leads to a number of human diseases, including cancer. We previously demonstrated that S100C/A11 was translocated to nuclei and inhibited DNA synthesis in human keratinocytes when exposed to high Ca2+. In the present study we examined the effects of synthetic partial peptides of S100C/A11 on human carcinoma cell lines. Only an N-terminal peptide with 19 amino acid residues (MAK19) showed cytotoxicity to the cell lines in dose- and time-dependent manners when introduced into cells by flanking the HIV-TAT protein transduction domain (TAT-MAK19). Pulse field electrophoresis revealed that DNA of the treated cells was partially degradated. Annexin V, a marker of cellular apoptosis, was detected in the cells treated with TAT-MAK19 by immunostaining and flow cytometry. The induction of apoptotic cell death was apparently independent of p53, p21WAF1/CIP1, and caspase activity, but treatment with TAT-MAK19 resulted in partial translocation of apoptosis-inducing factor (AIF) from the cytoplasm to nuclei. These results indicate that MAK19 induces apoptosis in human cell lines and may therefore lead to the establishment of a new molecular target for the treatment of human cancer.

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Abbreviations

AIF :

Apoptosis-inducing factor

z-VAD-fmk :

Benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone

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Acknowledgements

This work was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Dermatology, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikatachou, 700-8558, Okayama, Japan

    Eiichi Makino & Keiji Iwatsuki

  2. Department of Cell Biology, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikatachou, 700-8558, Okayama, Japan

    Masakiyo Sakaguchi & Nam-ho Huh

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  1. Eiichi Makino
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  2. Masakiyo Sakaguchi
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  3. Keiji Iwatsuki
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  4. Nam-ho Huh
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Correspondence to Nam-ho Huh.

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Makino, E., Sakaguchi, M., Iwatsuki, K. et al. Introduction of an N-terminal peptide of S100C/A11 into human cells induces apoptotic cell death. J Mol Med 82, 612–620 (2004). https://doi.org/10.1007/s00109-004-0560-1

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  • DOI: https://doi.org/10.1007/s00109-004-0560-1

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