Molecular and Cellular Biochemistry

, Volume 382, Issue 1–2, pp 1–17 | Cite as

Proteomic analysis of apoptotic and oncotic pancreatic acinar AR42J cells treated with caerulein

  • Jiangtao Chu
  • Hongliang Ji
  • Ming Lu
  • Zhituo Li
  • Xin Qiao
  • Bei Sun
  • Weihui Zhang
  • Dongbo XueEmail author


This study aims to determine the differentially expressed proteins in the pancreatic acinar cells undergoing apoptosis and oncosis stimulated with caerulein to explore different cell death process of the acinar cell. AR42J cells were treated with caerulein to induce cell model of acute pancreatitis. Cells that were undergoing apoptosis and oncosis were separated by flow cytometry. Then differentially expressed proteins in the two groups of separated cells were detected by shotgun liquid chromatography-tandem mass spectrometry. The results showed that 11 proteins were detected in both apoptosis group and oncosis group, 17 proteins were detected only in apoptosis group and 29 proteins were detected only in oncosis group. KEGG analysis showed that proteins detected only in apoptosis group were significantly enriched in 10 pathways, including ECM-receptor interaction, cell adhesion molecules, and proteins detected only in oncosis group were significantly enriched in three pathways, including endocytosis, base excision repair, and RNA degradation. These proteins we detected are helpful for us to understand the process of cell death in acute pancreatitis and may be useful for changing the death mode of pancreatic acinar cells, thus attenuating the severity of pancreatitis.


Proteomics Acute pancreatitis Apoptosis Oncosis 



This work was supported by the National Natural Science Foundation of China (30972907, 81070373); the National Science Foundation for Post-doctoral Scientists of China (Grant Number: 20090451020); Heilongjiang Postdoctoral Grant (Grant Number: LRB08-503); Science Foundation of the First Affiliated Hospital of Harbin Medical University (Grant Number: B08-003). Authors are most grateful to Prof. Wei Jiang and Dr. Wei Li (College of Bioinformatics Science and Technology, Harbin Medical University) for their help with sample bioinformatics analysis. We thank RCPA (Research Centre for Proteome Analysis, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, PR China) for providing the proteomics technology.

Conflict of interest

The authors declare that they have no competing interests.


  1. 1.
    Petersen OH, Tepikin AV, Gerasimenko JV, Gerasimenko OV, Sutton R, Criddle DN (2009) Fatty acids, alcohol and fatty acid ethylesters: toxic Ca2+ signal generation and pancreatitis. Cell Calcium 45:634–642CrossRefPubMedGoogle Scholar
  2. 2.
    Leung PS, Chan YC (2009) Role of oxidative stress in pancreatic inflammation. Antioxid Redox Signal 11:135–165CrossRefPubMedGoogle Scholar
  3. 3.
    Franco-Pons N, Gea-Sorlí S, Closa D (2010) Release of inflammatory mediators by adipose tissue during acute pancreatitis. J Pathol 221:175–182CrossRefPubMedGoogle Scholar
  4. 4.
    Golstein P, Kroemer G (2007) Cell death by necrosis: towards a molecular definition. Trends Biochem Sci 32:37–43CrossRefPubMedGoogle Scholar
  5. 5.
    Oiva J, Mustonen H, Kylänpää ML, Kyhälä L, Alanärä T, Aittomäki S, Siitonen S, Kemppainen E, Puolakkainen P, Repo H (2010) Patients with acute pancreatitis complicated by organ failure show highly aberrant monocyte signaling profiles assessed by phospho-specific flow cytometry. Crit Care Med 38:1702–1708CrossRefPubMedGoogle Scholar
  6. 6.
    Gorelick FS, Thrower E (2009) The acinar cell and early pancreatitis responses. Clin Gastroenterol Hepatol 7:S10–S14CrossRefPubMedGoogle Scholar
  7. 7.
    Xue DB, Zhang WH, Yun XG, Song C, Zheng B, Shi XY, Wang HY (2007) Regulating effects of arsenic trioxide on cell death pathways and inflammatory reactions of pancreatic acinar cells in rats. Chin Med J (Engl) 120:690–695Google Scholar
  8. 8.
    Pernice M, Dunn SR, Miard T, Dufour S, Dove S, Hoegh-Guldberg O (2011) Regulation of apoptotic mediators reveals dynamic responses to thermal stress in the reef building coral Acropora millepora. PLoS ONE 6:e16095CrossRefPubMedGoogle Scholar
  9. 9.
    Bredesen DE (2007) Key note lecture: toward a mechanistic taxonomy for cell death programs. Stroke 38:652–660CrossRefPubMedGoogle Scholar
  10. 10.
    Jugdutt BI, Idikio HA (2005) Apoptosis and oncosis in acute coronary syndromes: assessment and implications. Mol Cell Biochem 270:177–200CrossRefPubMedGoogle Scholar
  11. 11.
    Xue D, Zhang W, Liang T, Zhao S, Sun B, Sun D (2009) Effects of arsenic trioxide on the cerulein-induced AR42J cells and its gene regulation. Pancreas 38:e183–e189CrossRefPubMedGoogle Scholar
  12. 12.
    Jonas L, Mikkat U, Lehmann R, Schareck W, Walzel H, Schröder W, Lopp H, Püssa T, Toomik P (2003) Inhibitory effects of human and porcine alpha-amylase on CCK-8-stimulated lipase secretion of isolated rat pancreatic acini. Pancreatology 3:342–348CrossRefPubMedGoogle Scholar
  13. 13.
    Chaudhary P, Suryakumar G, Prasad R, Singh SN, Ali S, Ilavazhagan G (2012) Chronic hypobaric hypoxia mediated skeletal muscle atrophy: role of ubiquitin–proteasome pathway and calpains. Mol Cell Biochem 364:101–113CrossRefPubMedGoogle Scholar
  14. 14.
    Shcherbik N, Haines DS (2004) Ub on the move. J Cell Biochem 93:11–19CrossRefPubMedGoogle Scholar
  15. 15.
    Park SH, Bolender N, Eisele F, Kostova Z, Takeuchi J, Coffino P, Wolf DH (2007) The cytoplasmic Hsp70 chaperone machinery subjects misfolded and ER import incompetent proteins to degradation via the ubiquitin–proteasome system. Mol Biol Cell 18:153–165CrossRefPubMedGoogle Scholar
  16. 16.
    Löw P, Bussell K, Dawson SP, Billett MA, Mayer RJ, Reynolds SE (1997) Expression of a 26s proteasome ATPase subunit, MS73, in muscles that undergo developmentally programmed cell death, and its control by ecdysteroid hormones in the insect Manduca sexta. FEBS Lett 400:345–349CrossRefPubMedGoogle Scholar
  17. 17.
    Tan YY, Zhou HY, Wang ZQ, Chen SD (2008) Endoplasmic reticulum stress contributes to the cell death induced by UCH-L1 inhibitor. Mol Cell Biochem 318:109–115CrossRefPubMedGoogle Scholar
  18. 18.
    Kotamraju S, Tampo Y, Keszler A, Chitambar CR, Joseph J, Haas AL, Kalyanaraman B (2003) Nitric oxide inhibits H2O2-induced transferrin receptor-dependent apoptosis in endothelial cells: role of ubiquitin–proteasome pathway. Proc Natl Acad Sci USA 100:10653–10658CrossRefPubMedGoogle Scholar
  19. 19.
    Franchi L, McDonald C, Kanneganti TD, Amer A, Núñez G (2006) Nucleotide-binding oligomerization domain-like receptors: intracellular pattern recognition molecules for pathogen detection and host defense. J Immunol 177:3507–3513PubMedGoogle Scholar
  20. 20.
    Pétrilli V, Dostert C, Muruve DA, Tschopp J (2007) The inflammasome: a danger sensing complex triggering innate immunity. Curr Opin Immunol 19:615–622CrossRefPubMedGoogle Scholar
  21. 21.
    Kuida K, Lippke JA, Ku G, Harding MW, Livingston DJ, Su MS, Flavell RA (1995) Altered cytokine export and apoptosis in mice deficient in interleukin-1 beta converting enzyme. Science 267:2000–2003CrossRefPubMedGoogle Scholar
  22. 22.
    Brömme HJ, Holtz J (1996) Apoptosis in the heart: when and why? Mol Cell Biochem 163–164:261–275CrossRefPubMedGoogle Scholar
  23. 23.
    Kuida K, Lippke JA, Ku G, Harding MW, Livingston DJ, Su MS, Flavell RA (1997) Caspase-1 processes IFN-gamma-inducing factor and regulates LPS-induced IFN-gamma production. Nature 386:619–623CrossRefGoogle Scholar
  24. 24.
    Ghayur T, Banerjee S, Hugunin M, Butler D, Herzog L, Carter A, Quintal L, Sekut L, Talanian R, Paskind M, Wong W, Kamen R, Tracey D, Allen H (2000) An induced proximity model for NF-κB activation in the Nod1/RICK and RIP signaling pathways. J Biol Chem 275:27823–27831Google Scholar
  25. 25.
    Wang W, Luo BH (2010) Structural basis of integrin transmembrane activation. J Cell Biochem 109:447–452PubMedGoogle Scholar
  26. 26.
    Mercurio AM, Rabinovitz I (2001) Towards a mechanistic understanding of tumor invasion—lessons from the alpha6beta 4 integrin. Semin Cancer Biol 11:129–141CrossRefPubMedGoogle Scholar
  27. 27.
    Chung J, Mercurio AM (2004) Contributions of the alpha6 integrins to breast carcinoma survival and progression. Mol Cells 17:203–209PubMedGoogle Scholar
  28. 28.
    Yanagisawa K, Konishi H, Arima C, Tomida S, Takeuchi T, Shimada Y, Yatabe Y, Mitsudomi T, Osada H, Takahashi T (2010) Novel metastasis-related gene CIM functions in the regulation of multiple cellular stress–response pathways. Cancer Res 70:9949–9958CrossRefPubMedGoogle Scholar
  29. 29.
    Chen X, Sans MD, Strahler JR, Karnovsky A, Ernst SA, Michailidis G, Andrews PC, Williams JA (2010) Quantitative organellar proteomics analysis of rough endoplasmic reticulum from normal and acute pancreatitis rat pancreas. J Proteome Res 9:885–896CrossRefPubMedGoogle Scholar
  30. 30.
    Seixas C, Cruto T, Tavares A, Gaertig J, Soares H (2010) CCTalpha and CCTdelta chaperonin subunits are essential and required for cilia assembly and maintenance in tetrahymena. PLoS ONE 5:e10704CrossRefPubMedGoogle Scholar
  31. 31.
    Yu JH, Seo JY, Kim KH, Kim H (2008) Differentially expressed proteins in cerulean-stimulated pancreatic acinar cells: implication for acute pancreatitis. Int J Biochem Cell Biol 40:503–516CrossRefPubMedGoogle Scholar
  32. 32.
    Madureira PA, Hill R, Miller VA, Giacomantonio C, Lee PW, Waisman DM (2011) Annexin A2 is a novel cellular redox regulatory protein involved in tumorigenesis. Oncotarget 2:1075–1093PubMedGoogle Scholar
  33. 33.
    Cusick JK, Mustian A, Jacobs AT, Reyland ME (2012) Identification of PLSCR1 as a protein that interacts with RELT family members. Mol Cell Biochem 362:55–63CrossRefPubMedGoogle Scholar
  34. 34.
    Locksley RM, Killeen N, Lenardo MJ (2001) The TNF and TNF receptor superfamilies: integrating mammalian biology. Cell 104:487–501CrossRefPubMedGoogle Scholar
  35. 35.
    Cusick JK, Mustian A, Goldberg K, Reyland ME (2010) RELT induces cellular death in HEK 293 epithelial cells. Cell Immunol 261:1–8CrossRefPubMedGoogle Scholar
  36. 36.
    O’Konski MS, Pandol SJ (1990) Effects of caerulein on the apical cytoskeleton of the pancreatic acinar cell. J Clin Invest 86:1649–1657CrossRefPubMedGoogle Scholar
  37. 37.
    Jungermann J, Lerch MM, Weidenbach H, Lutz MP, Krüger B, Adler G (1995) Disassembly of rat pancreatic acinar cell cytoskeleton during supramaximal secretagogue stimulation. Am J Physiol 268:G328–G338PubMedGoogle Scholar
  38. 38.
    Ueda T, Takeyama Y, Kaneda K, Adachi M, Ohyanagi H, Saitoh Y (1992) Protective effect of a microtubule stabilizer taxol on caerulein-induced acute pancreatitis in rat. J Clin Invest 89:234–243CrossRefPubMedGoogle Scholar
  39. 39.
    Scaffidi P, Misteli T, Bianchi ME (2002) Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature 418:191–195CrossRefPubMedGoogle Scholar
  40. 40.
    Yang J, Huang C, Yang J, Jiang H, Ding J (2010) Statins attenuate high mobility group box-1 protein induced vascular endothelial activation: a key role for TLR4/NF-κB signaling pathway. Mol Cell Biochem 345:189–195CrossRefPubMedGoogle Scholar
  41. 41.
    Yuan H, Jin X, Sun J, Li F, Feng Q, Zhang C, Cao Y, Wang Y (2009) Protective effect of HMGB1 a box on organ injury of acute pancreatitis in mice. Pancreas 38:143–148CrossRefPubMedGoogle Scholar
  42. 42.
    Gong H, Zuliani P, Komuravelli A, Faeder JR, Clarke EM (2010) Analysis and verification of the HMGB1 signaling pathway. BMC Bioinformatics 11:S10CrossRefPubMedGoogle Scholar
  43. 43.
    Hao JJ, Zhu J, Zhou K, Smith N, Zhan X (2005) The coiled-coil domain is required for HS1 to bind to F-actin and activate Arp2/3 complex. J Biol Chem 280:37988–37994CrossRefPubMedGoogle Scholar
  44. 44.
    Takemoto Y, Sato M, Furuta M, Hashimoto Y (1996) Distinct binding patterns of HS1 to the Src SH2 and SH3 domains reflect possible mechanisms of recruitment and activation of downstream molecules. Int Immunol 8:1699–1705CrossRefPubMedGoogle Scholar
  45. 45.
    Uezu A, Horiuchi A, Kanda K, Kikuchi N, Umeda K, Tsujita K, Suetsugu S, Araki N, Yamamoto H, Takenawa T, Nakanishi H (2007) SGIP1α is an endocytic protein that directly interacts with phospholipids and Eps15. J Biol Chem 282:26481–26489CrossRefPubMedGoogle Scholar
  46. 46.
    Mellman I (1996) Endocytosis and molecular sorting. Annu Rev Cell Dev Biol 12:575–625CrossRefPubMedGoogle Scholar
  47. 47.
    Darshi M, Mendiola VL, Mackey MR, Murphy AN, Koller A, Perkins GA, Ellisman MH, Taylor SS (2011) Chchd3, an inner mitochondrial membrane protein, is essential for maintaining cristae integrity and mitochondrial function. J Biol Chem 286:2918–2932CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Jiangtao Chu
    • 1
  • Hongliang Ji
    • 1
  • Ming Lu
    • 2
  • Zhituo Li
    • 1
  • Xin Qiao
    • 2
  • Bei Sun
    • 1
  • Weihui Zhang
    • 1
  • Dongbo Xue
    • 1
    Email author
  1. 1.Department of General SurgeryThe First Affiliated Hospital of Harbin Medical UniversityNangang DistrictPeople’s Republic of China
  2. 2.Department of Surgery, David Geffen School of MedicineUniversity of Califonia at Los AngelesLos AngelesUSA

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