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Silencing of Hsp70 Intensifies 6-OHDA-Induced Apoptosis and Hsp90 Upregulation in PC12 Cells

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Abstract

By the current study, we tried to find out the interactive mechanisms enrolled by Hsp70 and Hsp90 following the 6-hydroxydopamine (6-OHDA)-induced oxidative stress. Of heat shock protein (Hsp) family, we have previously evaluated the effects of Hsp90 gene silencing on in vitro model of Parkinson’s disease and its influence on controlling the mechanisms of cell survival. Here, we extended our study to Hsp70 silencing short interfering RNA (siRNA) oligonucleotides, transfected into Pheochromocytoma (PC12) cells with/without exposure to 6-OHDA stress. In order to determine the probable effects of Hsp70 silencing on apoptotic factors, we assessed Bcl2/Bax ratio, nuclear level of PARP, and cleavage of caspase-3 under 6-OHDA stress condition. The results showed deteriorated effect of Hsp70 siRNA on apoptosis in cells exposed to only 6-OHDA. This is, at least in part, in consequence of upregulation of Hsp90, both at messenger RNA (mRNA) and protein levels. These data highlight the critical role of Hsp70 for cell survival under 6-OHDA stress condition. It could be a suggestive issue for supervision of caspase cascades by survival roles of Hsps as Hsp70 silencing resulted in apoptosis phenomenon. Convergence of Hsp70 anti-apoptotic and 6-OHDA pro-apoptotic pathways may explain intensified apoptosis following Hsp70 silencing. In addition, nuclear factor erythroid-2-related factor 2 (Nrf2), a transcription factor, has been previously studied in detoxification of oxidative stress. For this issue, we tried to elucidate Hsp70 silencing impact on Nrf2, which has been shown to regulate the transcription of Hsp70, unspecifically. Besides, our investigations revealed that Hsp70 siRNA did not affect the level of Nrf2 during 6-OHDA exposure. But, it is still a dealing question and other investigations are needed to have a comprehensive perception of Hsp family signaling functions.

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Acknowledgments

This work was supported financially by Iran National Science Foundation (grant number 90001703).

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Authors and Affiliations

  1. NeuroBiology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Payam Sadeghi, Fariba Khodagholi & Hadi Digaleh

  2. Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Payam Sadeghi, Fariba Khodagholi & Hadi Digaleh

  3. Department of Genetic and Molecular Biology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

    Behrang Alani, Rasoul Salehi & Hassan Korbekandi

  4. Department of Applied Cell Science, Faculty of Medicine, Kashan University of Medical Sciences, Kashan, Iran

    Behrang Alani

  5. Center for Cancer Stem Cell Research, Department of Medicine and Biosystemic Science and Graduate School of Medical Sciences, Kyushu University Hospital, Fukuoka, Japan

    Siamak Jabbarzadeh-Tabrizi

  6. Department of Neurology, Isfahan University of Medical Sciences, Isfahan, Iran

    Mohammad Zare

  7. Neuroscience Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

    Mohammad Zare

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  1. Behrang Alani
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  2. Rasoul Salehi
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  3. Payam Sadeghi
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  8. Hassan Korbekandi
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Correspondence to Behrang Alani.

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Alani, B., Salehi, R., Sadeghi, P. et al. Silencing of Hsp70 Intensifies 6-OHDA-Induced Apoptosis and Hsp90 Upregulation in PC12 Cells. J Mol Neurosci 55, 174–183 (2015). https://doi.org/10.1007/s12031-014-0298-3

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  • DOI: https://doi.org/10.1007/s12031-014-0298-3

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