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BNIP3 as a Regulator of Cisplatin-Induced Apoptosis

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Abstract

BNIP3 is a member of Bcl-2 protein family involved in regulation of various forms of cell death. However, its role in these processes remains unclear and varies depending on the type of cancer cells and environmental factors (pH, O2 level, etc.). Here, the role of BNIP3 in apoptosis regulation in lung adenocarcinoma cells was investigated. The suppressed expression of BNIP3 caused inhibition of oxygen consumption and stimulated production of the mitochondrial reactive oxygen species, suggesting the role of BNIP3 in induction of mitochondrial dysfunction and its potential involvement in regulation of cell death. Indeed, cytochrome c release in the cells with BNIP3 knockout and knockdown was higher than in the wild-type (WT) upon apoptosis stimulation by cisplatin. Moreover, suppression of BNIP3 expression led to the increase in the caspase-3 activity and, as a consequence, accumulation of the apoptotic marker – p89 fragment of poly(ADP-ribose)-polymerase (PARP) – as compared to WT cells. Analysis of the SubG1 population by flow cytometry confirmed the elevated level of apoptosis in the BNIP3 knockout cells. Pretreatment with the antioxidant Trolox did not affect cell death, indicating that it was independent on reactive oxygen species. These data show that BNIP3 is involved in maintaining normal functioning of mitochondria and, as a result, can regulate the mitochondrial pathway of cell death.

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Abbreviations

CCCP:

carbonyl cyanide-m-chlorophenylhydrazone

ETC:

electron-transport chain

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

mtROS:

mitochondrial reactive oxygen species

OCR:

oxygen consumption rate

PARP:

poly(ADP-ribose)-polymerase

siRNA:

small interfering RNA

WT:

wild-type

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Acknowledgements

The authors are thankful to Dr. Gelina Kopeina for providing valuable recommendations and feedback throughout this project.

Funding

This research was financially supported by the Russian Science Foundation (project no. 19-15-00125) and by the Russian Foundation for Basic Research (projects nos. 20-015-00105, 20-315-90019, and 19-015-00332), as well as by the Swedish (project no. 190345) and Stockholm (project no. 181301) Cancer Societies.

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Authors and Affiliations

  1. Faculty of Basic Medicine, Lomonosov Moscow State University, 119192, Moscow, Russia

    A. S. Gorbunova, T. V. Denisenko, M. A. Yapryntseva, A. D. Pivnyuk, V. G. Gogvadze & B. Zhivotovsky

  2. Skolkovo Institute of Science and Technology, Center of Life Sciences, 121205, Moscow, Russia

    T. A. Prikazchikova

  3. Institute of Environmental Medicine, Karolinska Institute, SE-17177, Stockholm, Sweden

    V. G. Gogvadze & B. Zhivotovsky

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  1. A. S. Gorbunova
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Correspondence to B. Zhivotovsky.

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The authors declare no conflicts of interest in financial or any other sphere. This article does not contain any studies with human participants or animals performed by any of the authors.

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Gorbunova, A.S., Denisenko, T.V., Yapryntseva, M.A. et al. BNIP3 as a Regulator of Cisplatin-Induced Apoptosis. Biochemistry Moscow 85, 1245–1253 (2020). https://doi.org/10.1134/S0006297920100120

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