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IL-1β induces ER stress in a JNK dependent manner that determines cell death in human pancreatic epithelial MIA PaCa-2 cells

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Abstract

The proinflammatory cytokine, IL-1β (Interleukin-1β) is a significant determinant of pancreatic apoptosis and cell death that are common characteristics during diabetes. Using human derived pancreatic MIA PaCa-2 cells, we describe one of the underlying molecular mechanisms behind this observation. Incubation of these cells with IL-1β at doses from 0.5 to 3.0 ng/ml caused significant cell death at 36 h. This was accompanied with marked increases in JNK and p38 phosphorylation together with increased levels of the endoplasmic reticulum (ER) stress markers, namely BiP, CHOP, GADD34, ATF4 and sXBP1. IL-1β also led to increased phosphorylation of eIF2α and all these events could be prevented by pretreatment with the JNK inhibitor, SP600125. A time course study indicated that while IL-1β mediated JNK phosphorylation was induced as early as 2 h of IL-1β treatment, induction of the ER stress markers was evident at later time points. IL-1β stimulated JNK phosphorylation was observed even in the presence of the ER stress inhibitor, 4-phenyl butyrate and the decrease in cell viability was significantly prevented in the presence of the JNK inhibitor. All these suggest that JNK activation is a pre-requisite for ER stress induction and cell death. Reports till date have consistently demonstrated JNK activation as a consequence of ER stress induction by IL-1β in the pancreas. We show here for the first time that the activation of JNK by IL-1β is a prelude to the subsequent induction of ER stress and cell death. These therefore suggest that the JNK-ER stress axis is critical in deciding the decreased survival status by IL-1β in MIA PaCa-2 cells.

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References

  1. Chandra J, Zhivotovsky B, Zaitsev S, Juntti-Berggren L, Berggren PO, Orrenius S (2001) Role of apoptosis in pancreatic beta-cell death in diabetes. Diabetes 50(Suppl 1):S44–S47

    Article  CAS  PubMed  Google Scholar 

  2. Wang Q, Zhang H, Zhao B, Fei H (2009) IL-1beta caused pancreatic beta-cells apoptosis is mediated in part by endoplasmic reticulum stress via the induction of endoplasmic reticulum Ca2+ release through the c-Jun N-terminal kinase pathway. Mol Cell Biochem 324:183–190

    Article  CAS  PubMed  Google Scholar 

  3. Oyadomari S, Takeda K, Takiguchi M, Gotoh T, Matsumoto M, Wada I, Akira S, Araki E, Mori M (2001) Nitric oxide-induced apoptosis in pancreatic beta cells is mediated by the endoplasmic reticulum stress pathway. Proc Natl Acad Sci USA 98:10845–10850

    Article  CAS  PubMed  Google Scholar 

  4. Akerfeldt MC, Howes J, Chan JY, Stevens VA, Boubenna N, McGuire HM, King C, Biden TJ, Laybutt DR (2008) Cytokine-induced beta-cell death is independent of endoplasmic reticulum stress signaling. Diabetes 57:3034–3044

    Article  CAS  PubMed  Google Scholar 

  5. Kharroubi I, Ladriere L, Cardozo AK, Dogusan Z, Cnop M, Eizirik DL (2004) Free fatty acids and cytokines induce pancreatic beta-cell apoptosis by different mechanisms: role of nuclear factor-kappaB and endoplasmic reticulum stress. Endocrinology 145:5087–5096

    Article  CAS  PubMed  Google Scholar 

  6. Ortis F, Cardozo AK, Crispim D, Storling J, Mandrup-Poulsen T, Eizirik DL (2006) Cytokine-induced proapoptotic gene expression in insulin-producing cells is related to rapid, sustained, and nonoscillatory nuclear factor-kappaB activation. Mol Endocrinol 20:1867–1879

    Article  CAS  PubMed  Google Scholar 

  7. Ortis F, Pirot P, Naamane N, Kreins AY, Rasschaert J, Moore F, Theatre E, Verhaeghe C, Magnusson NE, Chariot A, Orntoft TF, Eizirik DL (2008) Induction of nuclear factor-kappaB and its downstream genes by TNF-alpha and IL-1beta has a pro-apoptotic role in pancreatic beta cells. Diabetologia 51:1213–1225

    Article  CAS  PubMed  Google Scholar 

  8. Cardozo AK, Heimberg H, Heremans Y, Leeman R, Kutlu B, Kruhoffer M, Orntoft T, Eizirik DL (2001) A comprehensive analysis of cytokine-induced and nuclear factor-kappa B-dependent genes in primary rat pancreatic beta-cells. J Biol Chem 276:48879–48886

    Article  CAS  PubMed  Google Scholar 

  9. Eizirik DL, Mandrup-Poulsen T (2001) A choice of death—the signal-transduction of immune-mediated beta-cell apoptosis. Diabetologia 44:2115–2133

    Article  CAS  PubMed  Google Scholar 

  10. Rabinovitch A, Suarez-Pinzon WL (2003) Role of cytokines in the pathogenesis of autoimmune diabetes mellitus. Rev Endocr Metab Disord 4:291–299

    Article  CAS  PubMed  Google Scholar 

  11. Thomas HE, Darwiche R, Corbett JA, Kay TW (2002) Interleukin-1 plus gamma-interferon-induced pancreatic beta-cell dysfunction is mediated by beta-cell nitric oxide production. Diabetes 51:311–316

    Article  CAS  PubMed  Google Scholar 

  12. Corbett JA, McDaniel ML (1994) Reversibility of interleukin-1 beta-induced islet destruction and dysfunction by the inhibition of nitric oxide synthase. Biochem J 299:719–724

    CAS  PubMed  Google Scholar 

  13. Cnop M, Welsh N, Jonas JC, Jorns A, Lenzen S, Eizirik DL (2005) Mechanisms of pancreatic beta-cell death in type 1 and type 2 diabetes: many differences, few similarities. Diabetes 54(Suppl 2):S97–107

    Article  CAS  PubMed  Google Scholar 

  14. Cardozo AK, Ortis F, Storling J, Feng YM, Rasschaert J, Tonnesen M, Van EF, Mandrup-Poulsen T, Herchuelz A, Eizirik DL (2005) Cytokines downregulate the sarcoendoplasmic reticulum pump Ca2+ ATPase 2b and deplete endoplasmic reticulum Ca2+, leading to induction of endoplasmic reticulum stress in pancreatic beta-cells. Diabetes 54:452–461

    Article  CAS  PubMed  Google Scholar 

  15. Csala M, Banhegyi G, Benedetti A (2006) Endoplasmic reticulum: a metabolic compartment. FEBS Lett 580:2160–2165

    Article  CAS  PubMed  Google Scholar 

  16. Schroder M (2008) Endoplasmic reticulum stress responses. Cell Mol Life Sci 65:862–894

    Article  CAS  PubMed  Google Scholar 

  17. Ron D, Walter P (2007) Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 8:519–529

    Article  CAS  PubMed  Google Scholar 

  18. Wu J, Kaufman RJ (2006) From acute ER stress to physiological roles of the Unfolded Protein Response. Cell Death Differ 13:374–384

    Article  CAS  PubMed  Google Scholar 

  19. Laybutt DR, Preston AM, Akerfeldt MC, Kench JG, Busch AK, Biankin AV, Biden TJ (2007) Endoplasmic reticulum stress contributes to beta cell apoptosis in type 2 diabetes. Diabetologia 50:752–763

    Article  CAS  PubMed  Google Scholar 

  20. Huang CJ, Lin CY, Haataja L, Gurlo T, Butler AE, Rizza RA, Butler PC (2007) High expression rates of human islet amyloid polypeptide induce endoplasmic reticulum stress mediated beta-cell apoptosis, a characteristic of humans with type 2 but not type 1 diabetes. Diabetes 56:2016–2027

    Article  CAS  PubMed  Google Scholar 

  21. Harding HP, Zeng H, Zhang Y, Jungries R, Chung P, Plesken H, Sabatini DD, Ron D (2001) Diabetes mellitus and exocrine pancreatic dysfunction in perk−/− mice reveals a role for translational control in secretory cell survival. Mol Cell 7:1153–1163

    Article  CAS  PubMed  Google Scholar 

  22. Scheuner D, Song B, McEwen E, Liu C, Laybutt R, Gillespie P, Saunders T, Bonner-Weir S, Kaufman RJ (2001) Translational control is required for the unfolded protein response and in vivo glucose homeostasis. Mol Cell 7:1165–1176

    Article  CAS  PubMed  Google Scholar 

  23. Scheuner D, Vander MD, Song B, Flamez D, Creemers JW, Tsukamoto K, Ribick M, Schuit FC, Kaufman RJ (2005) Control of mRNA translation preserves endoplasmic reticulum function in beta cells and maintains glucose homeostasis. Nat Med 11:757–764

    Article  CAS  PubMed  Google Scholar 

  24. Zinszner H, Kuroda M, Wang X, Batchvarova N, Lightfoot RT, Remotti H, Stevens JL, Ron D (1998) CHOP is implicated in programmed cell death in response to impaired function of the endoplasmic reticulum. Genes Dev 12:982–995

    Article  CAS  PubMed  Google Scholar 

  25. Kaneto H, Nakatani Y, Kawamori D, Miyatsuka T, Matsuoka TA, Matsuhisa M, Yamasaki Y (2005) Role of oxidative stress, endoplasmic reticulum stress, and c-Jun N-terminal kinase in pancreatic beta-cell dysfunction and insulin resistance. Int J Biochem Cell Biol 37:1595–1608

    Article  CAS  PubMed  Google Scholar 

  26. Xu Z, Kukekov NV, Greene LA (2003) POSH acts as a scaffold for a multiprotein complex that mediates JNK activation in apoptosis. EMBO J 22:252–261

    Article  CAS  PubMed  Google Scholar 

  27. Xu Z, Kukekov NV, Greene LA (2005) Regulation of apoptotic c-Jun N-terminal kinase signaling by a stabilization-based feed-forward loop. Mol Cell Biol 25:9949–9959

    Article  CAS  PubMed  Google Scholar 

  28. Schroder M, Kaufman RJ (2005) The mammalian unfolded protein response. Annu Rev Biochem 74:739–789

    Article  PubMed  CAS  Google Scholar 

  29. Wang XZ, Lawson B, Brewer JW, Zinszner H, Sanjay A, Mi LJ, Boorstein R, Kreibich G, Hendershot LM, Ron D (1996) Signals from the stressed endoplasmic reticulum induce C/EBP-homologous protein (CHOP/GADD153). Mol Cell Biol 16:4273–4280

    CAS  PubMed  Google Scholar 

  30. Ma Y, Brewer JW, Diehl JA, Hendershot LM (2002) Two distinct stress signaling pathways converge upon the CHOP promoter during the mammalian unfolded protein response. J Mol Biol 318:1351–1365

    Article  CAS  PubMed  Google Scholar 

  31. Qiao HX, Hao CJ, Li Y, He X, Chen RS, Cui J, Xu ZH, Li W (2008) JNK activation mediates the apoptosis of xCT-deficient cells. Biochem Biophys Res Commun 370:584–588

    Article  CAS  PubMed  Google Scholar 

  32. Ichijo H, Nishida E, Irie K, ten Dijke P, Saitoh M, Moriguchi T, Takagi M, Matsumoto K, Miyazono K, Gotoh Y (1997) Induction of apoptosis by ASK1, a mammalian MAPKKK that activates SAPK/JNK and p38 signaling pathways. Science 275:90–94

    Article  CAS  PubMed  Google Scholar 

  33. Rutkowski DT, Arnold SM, Miller CN, Wu J, Li J, Gunnison KM, Mori K, Sadighi Akha AA, Raden D, Kaufman RJ (2006) Adaptation to ER stress is mediated by differential stabilities of pro-survival and pro-apoptotic mRNAs and proteins. PLoS Biol 4:e374

    Article  PubMed  CAS  Google Scholar 

  34. Nabokina SM, Ma TY, Said HM (2004) Mechanism and regulation of folate uptake by human pancreatic epithelial MIA PaCa-2 cells. Am J Physiol Cell Physiol 287:C142–C148

    Article  CAS  PubMed  Google Scholar 

  35. Wang F, Kumagai-Braesch M, Herrington MK, Larsson J, Permert J (2009) Increased lipid metabolism and cell turnover of MiaPaCa2 cells induced by high-fat diet in an orthotopic system. Metabolism 58:1131–1136

    Article  CAS  PubMed  Google Scholar 

  36. Madden ME, Heaton KM, Huff JK, Sarras MP Jr (1989) Comparative analysis of a human pancreatic undifferentiated cell line (MIA PaCa-2) to acinar and ductal cells. Pancreas 4:529–537

    Article  CAS  PubMed  Google Scholar 

  37. Kajino S, Suganuma M, Teranishi F, Takahashi N, Tetsuka T, Ohara H, Itoh M, Okamoto T (2000) Evidence that de novo protein synthesis is dispensable for anti-apoptotic effects of NF-kappaB. Oncogene 19:2233–2239

    Article  CAS  PubMed  Google Scholar 

  38. Khalooghi K, Hashemi S, Mehraban N, Amiri P, Bazzaz JT, Larijani B, Amoli MM (2009) In vitro modulation of TCF7L2 gene expression in human pancreatic cells. Mol Biol Rep 36:2329–2332

    Article  CAS  PubMed  Google Scholar 

  39. Gidez LI (1973) Effect of dietary fat on pancreatic lipase levels in the rat. J Lipid Res 14:169–177

    CAS  PubMed  Google Scholar 

  40. Eizirik DL, Welsh N, Hellerstrom C (1993) Predominance of stimulatory effects of interleukin-1 beta on isolated human pancreatic islets. J Clin Endocrinol Metab 76:399–403

    Article  CAS  PubMed  Google Scholar 

  41. Matsuda T, Ferreri K, Todorov I, Kuroda Y, Smith CV, Kandeel F, Mullen Y (2005) Silymarin protects pancreatic beta-cells against cytokine-mediated toxicity: implication of c-Jun NH2-terminal kinase and janus kinase/signal transducer and activator of transcription pathways. Endocrinology 146:175–185

    Article  CAS  PubMed  Google Scholar 

  42. Liao PC, Tan SK, Lieu CH, Jung HK (2008) Involvement of endoplasmic reticulum in paclitaxel-induced apoptosis. J Cell Biochem 104:1509–1523

    Article  CAS  PubMed  Google Scholar 

  43. Pandey AK, Bhardwaj V, Datta M (2009) Tumor necrosis factorα attenuates insulin action on phosphoenolpyruvate carboxykinase gene expression and gluconeogenesis by altering the cellular localization of Foxa2 in HepG2 cells. FEBS J 276:3757–3769

    Article  CAS  PubMed  Google Scholar 

  44. Choi SE, Lee YJ, Jang HJ, Lee KW, Kim YS, Jun HS, Kang SS, Chun J, Kang Y (2008) A chemical chaperone 4-PBA ameliorates palmitate-induced inhibition of glucose-stimulated insulin secretion (GSIS). Arch Biochem Biophys 475:109–114

    Article  CAS  PubMed  Google Scholar 

  45. Kutlu B, Cardozo AK, Darville MI, Kruhoffer M, Magnusson N, Orntoft T, Eizirik DL (2003) Discovery of gene networks regulating cytokine-induced dysfunction and apoptosis in insulin-producing INS-1 cells. Diabetes 52:2701–2719

    Article  CAS  PubMed  Google Scholar 

  46. Cardozo AK, Kruhoffer M, Leeman R, Orntoft T, Eizirik DL (2001) Identification of novel cytokine-induced genes in pancreatic beta-cells by high-density oligonucleotide arrays. Diabetes 50:909–920

    Article  CAS  PubMed  Google Scholar 

  47. Eizirik DL, Cardozo AK, Cnop M (2008) The role for endoplasmic reticulum stress in diabetes mellitus. Endocr Rev 29:42–61

    Article  CAS  PubMed  Google Scholar 

  48. Minden A, Karin M (1997) Regulation and function of the JNK subgroup of MAP kinases. Biochim Biophys Acta 1333:F85–104

    CAS  PubMed  Google Scholar 

  49. Hibi M, Lin A, Smeal T, Minden A, Karin M (1993) Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. Genes Dev 7:2135–2148

    Article  CAS  PubMed  Google Scholar 

  50. Liu J, Lin A (2005) Role of JNK activation in apoptosis: a double-edged sword. Cell Res 15:36–42

    Article  PubMed  Google Scholar 

  51. Kim S, Kim HS, Chung KW, Oh SH, Yun JW, Im SH, Lee MK, Kim KW, Lee MS (2007) Essential role for signal transducer and activator of transcription-1 in pancreatic beta-cell death and autoimmune type 1 diabetes of nonobese diabetic mice. Diabetes 56:2561–2568

    Article  CAS  PubMed  Google Scholar 

  52. Larsen CM, Faulenbach M, Vaag A, Volund A, Ehses JA, Seifert B, Mandrup-Poulsen T, Donath MY (2007) Interleukin-1-receptor antagonist in type 2 diabetes mellitus. N Engl J Med 356:1517–1526

    Article  CAS  PubMed  Google Scholar 

  53. Ehlers RA, Zhang Y, Hellmich MR, Evers BM (2000) Neurotensin-mediated activation of MAPK pathways and AP-1 binding in the human pancreatic cancer cell line, MIA PaCa-2. Biochem Biophys Res Commun 269:704–708

    Article  CAS  PubMed  Google Scholar 

  54. Hashimoto K, Farrow BJ, Evers BM (2004) Activation and role of MAP kinases in 15d-PGJ2-induced apoptosis in the human pancreatic cancer cell line MIA PaCa-2. Pancreas 28:153–159

    Article  CAS  PubMed  Google Scholar 

  55. Storling J, Zaitsev SV, Kapelioukh IL, Karlsen AE, Billestrup N, Berggren P, Mandrup-Poulsen T (2005) Calcium has a permissive role in IL-1β induce c-jun N-terminal kinase activation in insulin- secreting cells. Endocrinology 146:3026–3036

    Article  CAS  PubMed  Google Scholar 

  56. Oyadomari S, Araki E, Mori M (2002) Endoplasmic reticulum stress-mediated apoptosis in pancreatic β-cells. Apoptosis 7:335–345

    Article  CAS  PubMed  Google Scholar 

  57. Rutkowski DT, Kaufman RJ (2004) A trip to the ER: coping with stress. Trends Cell Biol 14:20–28

    Article  CAS  PubMed  Google Scholar 

  58. Kadowaki H, Nishitoh H, Ichijo H (2004) Survival and apoptosis signals in ER stress: the role of protein kinases. J Chem Neuroanat 28:93–100

    CAS  PubMed  Google Scholar 

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Acknowledgements

The authors are thankful to the Council of Scientific and Industrial Research (CSIR), New Delhi, India for financial support for carrying out this work (SIP006) and GV is thankful to CSIR for his fellowship (SIP006). We are thankful to Mr. Saurabh Vig for his help during the preparation of the illustrations being provided in this manuscript.

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Correspondence to Malabika Datta.

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Verma, G., Datta, M. IL-1β induces ER stress in a JNK dependent manner that determines cell death in human pancreatic epithelial MIA PaCa-2 cells. Apoptosis 15, 864–876 (2010). https://doi.org/10.1007/s10495-010-0498-4

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