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Essential role of caspases in epigallocatechin-3-gallate-mediated inhibition of nuclear factor kappaB and induction of apoptosis

Oncogene volume 23, pages 2507–2522 (2004)Cite this article

Abstract

Green tea constituent (−) epigallocatechin-3-gallate (EGCG) has shown remarkable cancer-preventive and some cancer-therapeutic effects. This is partially because of its ability to induce apoptosis in cancer cells without affecting normal cells. Previous studies from our laboratory have shown the involvement of NF-κB pathway in EGCG-mediated cell-cycle deregulation and apoptosis of human epidermoid carcinoma A431 cells. Here we show the essential role of caspases in EGCG-mediated inhibition of NF-κB and its subsequent apoptosis. Treatment of A431 cells with EGCG (10–40 μg/ml) resulted in dose-dependent inhibition of NF-κB/p65, induction of DNA breaks, cleavage of poly(ADP-ribose) polymerase (PARP) and morphological changes consistent with apoptosis. EGCG treatment of cells also resulted in significant activation of caspases, as shown by the dose- and time-dependent increase in DEVDase activity, and protein expression of caspase-3, -8 and -9. EGCG-mediated caspase activation induces proteolytic cleavage of NF-κB/p65 subunit, leading to the loss of transactivation domains, and driving the cells towards apoptosis. EGCG-mediated induction of apoptosis was significantly blocked by the caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (Z-VAD-FMK), and moderately blocked by the specific caspase-3 inhibitor Z-DEVD-FMK. Further, pretreatment of cells with Z-VAD-FMK was found to suppress the cleavage of NF-κB/p65 subunit, thereby increasing nuclear translocation, DNA binding and transcriptional activity, thus protecting the cells from EGCG-induced apoptosis. Taken together, these studies for the first time demonstrate that EGCG-mediated activation of caspases is critical, at least in part, for inhibition of NF-κB and subsequent apoptosis.

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Abbreviations

EGCG:

(−)-epigallocatechin-3-gallate

NF-κB:

nuclear factor-kappaB

PARP:

poly(ADP-ribose) polymerase

Z-VAD-FMK:

N-benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone

Z-DEVD-FMK:

CPP32/apopain inhibitor

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Acknowledgements

This work was supported by grants from United States Public Health Service (RO1CA 78809), American Institute for Cancer Research (00A030) and Department of Defense (DAMD 17-00-1-0527). Sanjay Gupta, PhD is grateful to the ‘Cancer Research Foundation of America’ and ‘The O-CHA (Tea) Pioneer Academic Research Grant Program’, Japan, for a partial support of funds.

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  1. Contributed equally to this work

Authors and Affiliations

  1. Department of Urology, Case Western Reserve University, Cleveland, 44106, OH, USA

    Sanjay Gupta

  2. Department of Environmental Health Sciences, Case Western Reserve University, Cleveland, 44106, OH, USA

    Kedar Hastak

  3. Department of Dermatology, Medical Sciences Center, 1300 University Avenue, University of Wisconsin, Madison, 53706, WI, USA

    Farrukh Afaq, Nihal Ahmad & Hasan Mukhtar

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Correspondence to Hasan Mukhtar.

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Gupta, S., Hastak, K., Afaq, F. et al. Essential role of caspases in epigallocatechin-3-gallate-mediated inhibition of nuclear factor kappaB and induction of apoptosis. Oncogene 23, 2507–2522 (2004). https://doi.org/10.1038/sj.onc.1207353

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