Download PDF
Horm Metab Res 2014; 46(08): 574-580
DOI: 10.1055/s-0034-1374588
Endocrine Research
© Georg Thieme Verlag KG Stuttgart · New York

Activation of eIF2α Signaling Cascade is Associated with Testosterone-Induced Cell Apoptosis in INS-1 Cells

Y. Cui
1   Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Key Laboratory for Assisted ­Reproduction and Reproductive Genetics, Shanghai, China
2   Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, China
3   National Research Center for Assisted Reproductive Technology and ­Reproductive Genetics, Jinan, China
4   The Key laboratory for Reproductive Endocrinology of Ministry of Education, Jinan, China
,
Z. Ma
2   Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, China
3   National Research Center for Assisted Reproductive Technology and ­Reproductive Genetics, Jinan, China
4   The Key laboratory for Reproductive Endocrinology of Ministry of Education, Jinan, China
,
H. Zhao
2   Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, China
3   National Research Center for Assisted Reproductive Technology and ­Reproductive Genetics, Jinan, China
4   The Key laboratory for Reproductive Endocrinology of Ministry of Education, Jinan, China
,
X. Chen
2   Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, China
3   National Research Center for Assisted Reproductive Technology and ­Reproductive Genetics, Jinan, China
4   The Key laboratory for Reproductive Endocrinology of Ministry of Education, Jinan, China
,
Y. Zhang
2   Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, China
3   National Research Center for Assisted Reproductive Technology and ­Reproductive Genetics, Jinan, China
4   The Key laboratory for Reproductive Endocrinology of Ministry of Education, Jinan, China
,
H. Guo
5   Qilu Hospital of Shandong University, Jinan, China
,
Y. Zhao
2   Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, China
3   National Research Center for Assisted Reproductive Technology and ­Reproductive Genetics, Jinan, China
4   The Key laboratory for Reproductive Endocrinology of Ministry of Education, Jinan, China
,
Z.-J. Chen
1   Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Key Laboratory for Assisted ­Reproduction and Reproductive Genetics, Shanghai, China
2   Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, China
3   National Research Center for Assisted Reproductive Technology and ­Reproductive Genetics, Jinan, China
4   The Key laboratory for Reproductive Endocrinology of Ministry of Education, Jinan, China
› Author Affiliations
Further Information

Publication History

received 29 November 2013

accepted 27 March 2014

Publication Date:
05 May 2014 (online)

Also available at
    Permissions and Reprints

Abstract

Hyperandrogenemia is associated with insulin resistance and type 2 diabetes in women with polycystic ovary syndrome raising the possibility that androgen receptor signaling pathway plays an important role in the development and progression of β-cell dysfunction. Testosterone is the major circulating androgen in women. In this study, we investigated the effect of testosterone on INS-1 cells to find whether excess androgen could produce endoplasmic reticulum (ER) stress thereby contributing to β-cell dysfunction. The role of testosterone in INS-1 cell apoptosis was detected by flow cytometry and electron microscopy. Expression of BIP, ATF4, and CHOP were assessed by RT-PCR and Western blot. Testosterone/AR could not only initiate cell apoptosis but also induce the activation of eukaryotic initiation factor 2 alpha (eIF2α) cascades in INS-1 cells. Treatment of ER stress inhibitor or flutamide (AR inhibitor) could inhibit testosterone-induced cell apoptosis and CHOP expression. These results suggest that testosterone/AR pathway caused INS-1 cell apoptosis was at least in part through eIF2α/CHOP cascades.

Key words

INS-1 cells - testosterone - apoptosis - CHOP

Supporting Information

    for this article is available online at http://www.thieme-connect.de/ejournals/toc/hmr
  • Supporting Information
 

  • References

  • 1 Liu S, Navarro G, Mauvais-Jarvis F. Androgen excess produces systemic oxidative stress and predisposes to beta-cell failure in female mice. PloS One 2010; 24: e11302
    PubMedSearch in Google Scholar
  • 2 Diamond MP, Grainger D, Diamond MC, Sherwin RS, Defronzo RA. Effects of methyltestosterone on insulin secretion and sensitivity in women. J Clin Endocrinol Metab 1998; 83: 4420-4425
    CrossrefPubMedSearch in Google Scholar
  • 3 Goodarzi MO, Erickson S, Port SC, Jennich RI, Korenman SG. Beta-cell function: a key pathological determinant in polycystic ovary syndrome. J Clin Endocrinol Metab 2005; 90: 310-315
    CrossrefPubMedSearch in Google Scholar
  • 4 Rhodes CJ. Type 2 diabetes- a matter of beta-cell life and death?. Science 2005; 307: 380-384
    CrossrefPubMedSearch in Google Scholar
  • 5 Marciniak SJ, Ron D. Endoplasmic reticulum stress signaling in disease. Physiol Rev 2006; 86: 1133-1149
    CrossrefPubMedSearch in Google Scholar
  • 6 Dai R, Yan D, Li J, Chen S, Liu Y, Chen R, Duan C, Wei M, Li H, He T. Activation of PKR/eIF2α signaling cascade is associated with dihydrotestosterone-induced cell cycle arrest and apoptosis in human liver cells. J Cell Biochem 2012; 113: 1800-1808
    PubMedSearch in Google Scholar
  • 7 Cui Y, Ma J, Shi Y, Huan Q, Guo H, Zhao Y, Chen ZJ. Positive and negative feedback regulation in the production and secretion of insulin from INS-1 cells by testosterone. Horm Metab Res 2011; 43: 1-8
    Thieme ConnectPubMedSearch in Google Scholar
  • 8 Ye J, Rawson RB, Komuro R, Chen X, Davé UP, Prywes R, Brown MS, Goldstein JL. ER stress induces cleavage of membrane-bound ATF6 by the same proteases that process SREBPs. Mol Cell 2000; 6: 1355-1364
    CrossrefPubMedSearch in Google Scholar
  • 9 Yoshida H, Okada T, Haze K, Yanagi H, Yura T, Negishi M, Mori K. ATF6 activated by proteolysis binds in the presence of NF-Y (CBF) directly to the cis-acting element responsible for the mammalian unfolded protein response. Mol Cell Biol 2000; 20: 6755-6767
    CrossrefPubMedSearch in Google Scholar
  • 10 Haze K, Yoshida H, Yanagi H, Yura T, Mori K. Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress. Mol Biol Cell 1999; 10: 3787-3799
    CrossrefPubMedSearch in Google Scholar
  • 11 O’Meara NM, Blackman JD, Ehrmann DA, Barnes RB, Jaspan JB, Rosenfield RL, Polonsky KS. Defects in beta-cell function in functional ovarian hyperandrogenism. J Clin Endocrinol Metab 1993; 76: 1241-1247
    PubMedSearch in Google Scholar
  • 12 Azziz R. Controversy in clinical endocrinology: diagnosis of polycystic ovarian syndrome: the Rotterdam criteria are premature. J Clin Endocrinol Metab 2006; 91: 781-785
    CrossrefPubMedSearch in Google Scholar
  • 13 Azziz R, Carmina E, Dewailly D, Diamanti-Kandarakis E, Escobar-Morreale HF, Futterweit W, Janssen OE, Legro RS, Norman RJ, Taylor AE, Witchel SF. Androgen Excess Society . Positions statement: criteria for defining polycystic ovary syndrome as a predominantly hyperandrogenic syndrome: an Androgen Excess Society guideline. J Clin Endocrinol Metab 2006; 91: 4237-4245
    Search in Google Scholar
  • 14 Paik SG, Michelis MA, Kim YT, Shin S. Induction of insulin-dependent diabetes by streptozotocin. Inhibition by estrogens and potentiation by androgens. Diabetes 1982; 31: 724-729
    CrossrefPubMedSearch in Google Scholar
  • 15 Arora DK, Machhadieh B, Matti A, Wadzinski BE, Ramanadham S, Kowluru A. High glucose exposure promotes activation of protein phosphatase 2A in rodent islets and INS-1 832/13 β-Cells by increasing the posttranslational carboxylmethylation of its catalytic subunit. Endocrinology 2014; 155: 380-391
    CrossrefPubMedSearch in Google Scholar
  • 16 Tang C, Koulajian K, Schuiki I, Zhang L, Desai T, Ivovic A, Wang P, Robson-Doucette C, Wheeler MB, Minassian B, Volchuk A, Giacca A. Glucose-induced beta cell dysfunction in vivo in rats: link between oxidative stress and endoplasmic reticulum stress. Diabetologia 2012; 55: 1366-1379
    CrossrefPubMedSearch in Google Scholar
  • 17 Hu P, Han Z, Couvillon AD, Kaufman RJ, Exton JH. Autocrine tumor necrosis factor alpha links endoplasmic reticulum stress to the membrane death receptor pathway through IRE1 alpha-mediated NF-kappaB activation and down-regulation of TRAF2 expression. Mol Cell Biol 2006; 26: 3071-3084
    CrossrefPubMedSearch in Google Scholar
  • 18 Urano F, Wang X, Bertolotti A, Zhang Y, Chung P, Harding HP, Ron D. Coupling of stress in the ER to activation of JNK protein kinases by transmembrane protein kinase IRE1. Science 2000; 287: 664-666
    CrossrefPubMedSearch in Google Scholar
  • 19 Laybutt DR, Preston AM, Akerfeldt MC, Kench JG, Busch AK, Biankin AV, Biden TJ. Endoplasmic reticulum stress contributes to beta cell apoptosis in type 2 diabetes. Diabetologia 2007; 50: 752-763
    CrossrefPubMedSearch in Google Scholar
  • 20 Araki E, Oyadomari S, Mori M. Impact of endoplasmic reticulum stress pathway on pancreatic beta-cells and diabetes mellitus. Exp Biol Med (Maywood) 2003; 228: 1213-1217
    PubMedSearch in Google Scholar
  • 21 Karaskov E, Scott C, Zhang L, Teodoro T, Ravazzola M, Volchuk A. Chronic palmitate but not oleate exposure induces endoplasmic reticulum stress, which may contribute to INS-1 pancreatic beta-cell apoptosis. Endocrinology 2006; 147: 3398-3407
    CrossrefPubMedSearch in Google Scholar
  • 22 Marchetti P, Bugliani M, Lupi R, Marselli L, Masini M, Boggi U, Filipponi F, Weir GC, Eizirik DL, Cnop M. The endoplasmic reticulum in pancreatic beta cells of type 2 diabetes patients. Diabetologia 2007; 50: 2486-2494
    CrossrefPubMedSearch in Google Scholar
  • 23 Oyadomari S, Takeda K, Takiguchi M, Gotoh T, Matsumoto M, Wada I, Akira S, Akaki E, Mori M. Nitric oxide-induced apoptosis in pancreatic beta cells is mediated by the endoplasmic reticulum stress pathway. Proc Natl Acad Sci USA 2001; 98: 10845-10850
    CrossrefPubMedSearch in Google Scholar
  • 24 Kharroubi I, Ladrière L, Cardozo AK, Dogusan Z, Cnop M, Eizirik DL. Free fatty acids and cytokines induce pancreatic beta-cell apoptosis by different mechanisms: role of nuclear factor-kappaB and endoplasmic reticulum stress. Endocrinology 2004; 145: 5087-5096
    CrossrefPubMedSearch in Google Scholar
  • 25 Iyer S, Korada M, Rainbow L, Kirk J, Brown RM, Shaw N, Barrett TG. Wolcott-Rallison syndrome: a clinical and genetic study of three children, novel mutation in EIF2AK3 and a review of the literature. Acta Paediatr 2004; 93: 1195-1201
    CrossrefPubMedSearch in Google Scholar
  • 26 Oyadomari S, Koizumi A, Takeda K, Gotoh T, Akira S, Araki E, Mori M. Targeted disruption of the Chop gene delays endoplasmic reticulum stress-mediated diabetes. J Clin Invest 2002; 109: 525-532
    CrossrefPubMedSearch in Google Scholar
  • 27 Osman AA, Saito M, Makepeace C, Permutt MA, Schlesinger P, Mueckler M. Wolframin expression induces novel ion channel activity in endoplasmic reticulum membranes and increases intracellular calcium. J Biol Chem 2003; 278: 52755-52762
    CrossrefPubMedSearch in Google Scholar
  • 28 Schröder M, Kaufman RJ. ER stress and the unfolded protein response. Mutat Res 2005; 569: 29-63
    CrossrefPubMedSearch in Google Scholar
  • 29 Junger E, Herberg L, Jeruschke K, Leiter EH. The diabetes-prone NZO/HI strain. II. Pancreatic immunopathology. Lab Invest 2002; 82: 843-853
    CrossrefPubMedSearch in Google Scholar