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Nifedipine prevents etoposide-induced caspase-3 activation, prenyl transferase degradation and loss in cell viability in pancreatic β-cells

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Abstract

Emerging evidence implicates novel roles for post-translational prenylation (i.e., farnesylation and geranylgeranylation) of various signaling proteins in a variety of cellular functions including hormone secretion, survival and apoptosis. In the context of cellular apoptosis, it has been shown previously that caspase-3 activation, a hallmark of mitochondrial dysregulation, promotes hydrolysis of several key cellular proteins. We report herein that exposure of insulin-secreting INS 832/13 cells or normal rat islets to etoposide leads to significant activation of caspase-3 and subsequent degradation of the common α-subunit of farnesyl/geranylgeranyl transferases (FTase/GGTase). Furthermore, the above stated signaling steps were prevented by Z-DEVD-FMK, a known inhibitor of caspase-3. In addition, treatment of cell lysates with recombinant caspase-3 also caused FTase/GGTase α-subunit degradation. Moreover, nifedipine, a calcium channel blocker, markedly attenuated etoposide-induced caspase-3 activation, FTase/GGTase α-subunit degradation in INS 832/13 cells and normal rat islets. Further, nifedipine significantly restored etoposide-induced loss in metabolic cell viability in INS 832/13 cells. Based on these findings, we conclude that etoposide induces loss in cell viability by inducing mitochondrial dysfunction, caspase-3 activation and degradation of FTase/GGTase α-subunit. Potential significance of these findings in the context of protein prenylation and β-cell survival are discussed.

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Abbreviations

CSP3:

Caspase-3

ET:

Etoposide

FTase α:

Farnesyltransferase α subunit

GGTase α:

Geranylgeranyltransferase α subunit

Δ FTase/GGTase:

Hydrolytic product of FTase/GGTase α-subunit

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

Nif:

Nifedipine

References

  1. Kowluru A (2010) Small G proteins in islet beta-cell function. Endocr Rev 31:52–78

    Article  PubMed  CAS  Google Scholar 

  2. Robertson RP, Seaquist ER, Walseth TF (1991) G proteins and modulation of insulin secretion. Diabetes 40:1–6

    Article  PubMed  CAS  Google Scholar 

  3. Cox AD, Der CJ (1992) Protein prenylation: more than just glue? Curr Opin Cell Biol 4:1008–1016

    Article  PubMed  CAS  Google Scholar 

  4. Metz SA, Rabaglia ME, Stock JB, Kowluru A (1993) Modulation of insulin secretion from normal rat islets by inhibitors of the post-translational modifications of GTP-binding proteins. Biochem J. 295:31–40

    PubMed  CAS  Google Scholar 

  5. Kowluru A (2003) Regulatory roles for small G proteins in the pancreatic beta-cell: lessons from models of impaired insulin secretion. Am J Physiol: Endocrinol Metab 285:E669–E684

    CAS  Google Scholar 

  6. Casey PJ, Seabra MC (1996) Protein prenyltransferases. J Biol Chem 271:5289–5292

    Article  PubMed  CAS  Google Scholar 

  7. Lane KT, Beese LS (2006) Thematic review series: lipid posttranslational modifications. Structural biology of protein farnesyltransferase and geranylgeranyltransferase type I. J Lipid Res 47:681–699

    Article  PubMed  CAS  Google Scholar 

  8. Amin R, Chen HQ, Tannous M, Gibbs R, Kowluru A (2002) Inhibition of glucose- and calcium-induced insulin secretion from betaTC3 cells by novel inhibitors of protein isoprenylation. J Pharmacol Exp Ther 303:82–88

    Article  PubMed  CAS  Google Scholar 

  9. Veluthakal R, Kaur H, Goalstone M, Kowluru A (2007) Dominant-negative alpha-subunit of farnesyl- and geranyltransferase inhibits glucose-stimulated, but not KCl-stimulated, insulin secretion in INS 832/13 cells. Diabetes 56:204–210

    Article  PubMed  CAS  Google Scholar 

  10. Kowluru A (2008) Protein prenylation in glucose-induced insulin secretion from the pancreatic islet beta-cell: a perspective. J Cell Mol Med 12:164–173

    Article  PubMed  CAS  Google Scholar 

  11. Kowluru A, Veluthakal R, Rhodes CJ, Kamath V, Syed I, Koch BJ (2010) Protein farnesylation-dependent Raf/extracellular signal-related kinase signaling links to cytoskeletal remodeling to facilitate glucose-induced insulin secretion in pancreatic beta-cells. Diabetes 59:967–977

    Article  PubMed  CAS  Google Scholar 

  12. Goalstone M, Kamath V, Kowluru A (2010) Glucose activates prenyltransferases in pancreatic islet beta-cells. Biochem Biophys Res Commun 391:895–898

    Article  PubMed  CAS  Google Scholar 

  13. Kim KW, Chung HH, Chung CW et al (2001) Inactivation of farnesyltransferase and geranylgeranyltransferase I by caspase-3: cleavage of the common alpha subunit during apoptosis. Oncogene 20:358–366

    Article  PubMed  CAS  Google Scholar 

  14. Collier JJ, Fueger PT, Hohmeier HE, Newgard CB (2006) Pro- and antiapoptotic proteins regulate apoptosis but do not protect against cytokine-mediated cytotoxicity in rat islets and beta-cell lines. Diabetes 55:1398–1406

    Article  PubMed  CAS  Google Scholar 

  15. Duchen MR (2000) Mitochondria and calcium: from cell signalling to cell death. J Physiol 529:57–68

    Article  PubMed  CAS  Google Scholar 

  16. Sharma AK, Rohrer B (2004) Calcium-induced calpain mediates apoptosis via caspase-3 in a mouse photoreceptor cell line. J Biol Chem 279:35564–35572

    Article  PubMed  CAS  Google Scholar 

  17. Tripathi A, Chaube SK (2012) High cytosolic free calcium level signals apoptosis through mitochondria-caspase mediated pathway in rat eggs cultured in vitro. Apoptosis 17:439–448

    Article  PubMed  CAS  Google Scholar 

  18. Smith MA, Schnellmann RG (2012) Calpains mitochondria and apoptosis. Cardiovasc Res. doi:10.1093/cvr/cvs163

    PubMed  Google Scholar 

  19. Robertson JD, Gogvadze V, Zhivotovsky B, Orrenius S (2000) Distinct pathways for stimulation of cytochrome c release by etoposide. J Biol Chem 275:32438–32443

    Article  PubMed  CAS  Google Scholar 

  20. Parihar A, Parihar MS, Ghafourifar P (2008) Significance of mitochondrial calcium and nitric oxide for apoptosis of human breast cancer cells induced by tamoxifen and etoposide. Int J Mol Med 21:317–324

    PubMed  CAS  Google Scholar 

  21. Sorkin EM, Clissold SP, Brogden RN (1985) Nifedipine. A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic efficacy, in ischaemic heart disease, hypertension and related cardiovascular disorders. Drugs. 30:182–274

    Article  PubMed  CAS  Google Scholar 

  22. Bean BP (1989) Classes of calcium channels in vertebrate cells. Annu Rev Physiol 51:367–384

    Article  PubMed  CAS  Google Scholar 

  23. Sane KM, Mynderse M, LaLonde DT et al (2010) A novel geranylgeranyl transferase inhibitor in combination with lovastatin inhibits proliferation and induces autophagy in STS-26T MPNST cells. J Pharmacol Exp Ther 333:23–33

    Article  PubMed  CAS  Google Scholar 

  24. Geryk-Hall M, Yang Y, Hughes DP (2010) Driven to death: inhibition of farnesylation increases ras activity in osteosarcoma and promotes growth arrest and cell death. Mol Cancer Ther 9:1111–1119

    Article  PubMed  CAS  Google Scholar 

  25. Li G, Segu VB, Rabaglia ME, Luo RH, Kowluru A, Metz SA (1998) Prolonged depletion of guanosine triphosphate induces death of insulin-secreting cells by apoptosis. Endocrinology 139:3752–3762

    Article  PubMed  CAS  Google Scholar 

  26. Wang Y, Gao L, Li Y, Chen H, Sun Z (2011) Nifedipine protects INS-1 beta-cell from high glucose-induced ER Stress and apoptosis. Int J Mol Sci 12:7569–7580

    Article  PubMed  CAS  Google Scholar 

  27. Xu G, Chen J, Jing G, Shalev A (2012) Preventing β-cell loss and diabetes with calcium channel blockers. Diabetes 61:848–856

    Article  PubMed  CAS  Google Scholar 

  28. Kyathanahalli CN, Kowluru A (2011) A farnesylated G-protein suppresses Akt phosphorylation in INS 832/13 cells and normal rat islets: regulation by pertussis toxin and PGE(2). Biochem Pharmacol 81:1237–1247

    Article  PubMed  CAS  Google Scholar 

  29. Lutz RJ, Trujillo MA, Denham KS, Wenger L, Sinensky M (1992) Nucleoplasmic localization of prelamin A: implications for prenylation-dependent lamin A assembly into the nuclear lamina. Proc Natl Acad Sci USA 89:3000–3004

    Article  PubMed  CAS  Google Scholar 

  30. Prokocimer M, Davidovich M, Nissim-Rafinia M et al (2009) Nuclear lamins: key regulators of nuclear structure and activities. J Cell Mol Med 13:1059–1085

    Article  PubMed  CAS  Google Scholar 

  31. Chang SY, Hudon-Miller SE, Yang SH et al (2012) Inhibitors of protein geranylgeranyltransferase-I lead to prelamin A accumulation in cells by inhibiting ZMPSTE24. J Lipid Res 53:1176–1182

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This research was supported in part by a Merit Review award [to AK; 1BX000469] from the Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development Biomedical Laboratory Research and Development], and by the NIH/NIDDK [RO1 DK74921]. AK is also the recipient of a Senior Research Career Scientist Award from the Department of VA. The authors would like to thank Prof. Chris Newgard for INS 832/13 cells. We thank Dr. Wasanthi Subasinghe for excellent technical assistance in these studies.

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Authors and Affiliations

  1. Beta-Cell Biochemistry Laboratory, John D. Dingell VA Medical Center, 4646 John R, Detroit, MI, 48201, USA

    Daleep K. Arora, Abiy M. Mohammed & Anjaneyulu Kowluru

  2. Department of Pharmaceutical Sciences, Eugene Applebaum College of Pharmacy and Health Sciences, Wayne State University, Detroit, MI, 48201, USA

    Daleep K. Arora, Abiy M. Mohammed & Anjaneyulu Kowluru

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  1. Daleep K. Arora
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  2. Abiy M. Mohammed
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Correspondence to Anjaneyulu Kowluru.

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Arora, D.K., Mohammed, A.M. & Kowluru, A. Nifedipine prevents etoposide-induced caspase-3 activation, prenyl transferase degradation and loss in cell viability in pancreatic β-cells. Apoptosis 18, 1–8 (2013). https://doi.org/10.1007/s10495-012-0763-9

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