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Effect of PKCα expression on Bcl-2 phosphorylation and cell death by hypericin

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Abstract

In order to explain the contribution of the protein kinase Cα (PKCα) in apoptosis induced by photo-activation of hypericin (Hyp), a small interfering RNA was used for post-transcriptional silencing of pkcα gene expression. We have evaluated the influence of Hyp photo-activation on cell death in non-transfected and transfected (PKCα−) human glioma cells (U-87 MG). No significant differences were detected in cell survival between non-transfected and transfected PKCα− cells. However, the type of cell death was notably affected by silencing the pkcα gene. Photo-activation of Hyp strongly induced apoptosis in non-transfected cells, but the level of necrotic cells in transfected PKCα− cells increased significantly. The differences in cell death after Hyp photo-activation are demonstrated by changes in: (i) reactive oxygen species production, (ii) Bcl-2 phosphorylation on Ser70 (pBcl-2(Ser70)), (iii) cellular distributions of pBcl-2(Ser70) and (iv) cellular distribution of endogenous anti-oxidant glutathione and its co-localization with mitochondria. In summary, we suggest that post-transcriptional silencing of the pkcα gene and the related decrease of PKCα level considerably affects the anti-apoptotic function and the anti-oxidant function of Bcl-2. This implies that PKCα, as Bcl-2 kinase, indirectly protects U-87 MG cells against oxidative stress and subsequent cell death.

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Acknowledgments

This work was supported by: the FP7 EU project CELIM 316310, the project of the Slovak Res. and Dev. Agency APVV-0242-11, the project of the Agency of the Ministry of Education of Slovak (Republic for the Structural funds of the European Union: Doktorand (ITMS code: 26110230013) and KVARK (ITMS code: 26110230084) and by the International Program for Scientific Cooperation (PICS N°5398) from the CNRS. This work forms a part of the co-tutoring doctoral study of J. J. (P. J. Safarik University and UPMC).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Biophysics, Faculty of Science, University of Pavol Jozef Safarik, Jesenna 5, 041 54, Kosice, Slovak Republic

    Jaroslava Joniova, Matus Misuth, Pavol Miskovsky & Zuzana Nadova

  2. Laboratoire Jean Perrin LJP, CNRS/UPMC Univ Paris 06, FRE 3231, 75005, Paris, France

    Jaroslava Joniova & Franck Sureau

  3. Centre for Interdisciplinary Biosciences, University of Pavol Jozef Safarik, Kosice, Slovak Republic

    Pavol Miskovsky & Zuzana Nadova

Authors
  1. Jaroslava Joniova
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  2. Matus Misuth
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  3. Franck Sureau
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  4. Pavol Miskovsky
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  5. Zuzana Nadova
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Correspondence to Zuzana Nadova.

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Joniova, J., Misuth, M., Sureau, F. et al. Effect of PKCα expression on Bcl-2 phosphorylation and cell death by hypericin. Apoptosis 19, 1779–1792 (2014). https://doi.org/10.1007/s10495-014-1043-7

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