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Nicotine/cigarette smoke promotes metastasis of pancreatic cancer through α7nAChR-mediated MUC4 upregulation

Oncogene volume 32, pages 1384–1395 (2013)Cite this article

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Abstract

Despite evidence that long-term smoking is the leading risk factor for pancreatic malignancies, the underlying mechanism(s) for cigarette-smoke (CS)-induced pancreatic cancer (PC) pathogenesis has not been well established. Our previous studies revealed an aberrant expression of the MUC4 mucin in PC as compared with the normal pancreas, and its association with cancer progression and metastasis. Interestingly, here we explore a potential link between MUC4 expression and smoking-mediated PC pathogenesis and report that both cigarette smoke extract and nicotine, which is the major component of CS, significantly upregulates MUC4 in PC cells. This nicotine-mediated MUC4 overexpression was via the α7 subunit of nicotinic acetylcholine receptor (nAChR) stimulation and subsequent activation of the JAK2/STAT3 downstream signaling cascade in cooperation with the MEK/ERK1/2 pathway; this effect was blocked by the α7nAChR antagonists, α-bungarotoxin and mecamylamine, and by specific siRNA-mediated STAT3 inhibition. In addition, we demonstrated that nicotine-mediated MUC4 upregulation promotes the PC cell migration through the activation of the downstream effectors, such as HER2, c-Src and FAK; this effect was attenuated by shRNA-mediated MUC4 abrogation, further implying that these nicotine-mediated pathological effects on PC cells are MUC4 dependent. Furthermore, the in vivo studies showed a marked increase in the mean pancreatic tumor weight (low dose (100 mg/m3 total suspended particulate (TSP)), P=0.014; high dose (247 mg/m3 TSP), P=0.02) and significant tumor metastasis to various distant organs in the CS-exposed mice, orthotopically implanted with luciferase-transfected PC cells, as compared with the sham controls. Moreover, the CS-exposed mice had elevated levels of serum cotinine (low dose, 155.88±35.96 ng/ml; high dose, 216.25±29.95 ng/ml) and increased MUC4, α7nAChR and pSTAT3 expression in the pancreatic tumor tissues. Altogether, our findings revealed for the first time that CS upregulates the MUC4 mucin in PC via the α7nAChR/JAK2/STAT3 downstream signaling cascade, thereby promoting metastasis of PC.

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References

  1. Boyle P, Maisonneuve P, Bueno de MB, Ghadirian P, Howe GR, Zatonski W et alCigarette smoking and pancreas cancer: a case control study of the search programme of the IARC. Int J Cancer 1996; 67: 63–71.

    Article  CAS  PubMed  Google Scholar 

  2. Muscat JE, Stellman SD, Hoffmann D, Wynder EL . Smoking and pancreatic cancer in men and women. Cancer Epidemiol Biomarkers Prev 1997; 6: 15–19.

    CAS  PubMed  Google Scholar 

  3. Fuchs CS, Colditz GA, Stampfer MJ, Giovannucci EL, Hunter DJ, Rimm EB et alA prospective study of cigarette smoking and the risk of pancreatic cancer. Arch Intern Med 1996; 156: 2255–2260.

    Article  CAS  PubMed  Google Scholar 

  4. Wittel UA, Singh AP, Henley BJ, Andrianifahanana M, Akhter MP, Cullen DM et alCigarette smoke-induced differential expression of the genes involved in exocrine function of the rat pancreas. Pancreas 2006; 33: 364–370.

    Article  CAS  PubMed  Google Scholar 

  5. Malfertheiner P, Schutte K . Smoking—a trigger for chronic inflammation and cancer development in the pancreas. Am J Gastroenterol 2006; 101: 160–162.

    Article  CAS  PubMed  Google Scholar 

  6. Heeschen C, Jang JJ, Weis M, Pathak A, Kaji S, Hu RS et alNicotine stimulates angiogenesis and promotes tumor growth and atherosclerosis. Nat Med 2001; 7: 833–839.

    Article  CAS  PubMed  Google Scholar 

  7. Heeschen C, Weis M, Aicher A, Dimmeler S, Cooke JP . A novel angiogenic pathway mediated by non-neuronal nicotinic acetylcholine receptors. J Clin Invest 2002; 110: 527–536.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Wittel UA, Pandey KK, Andrianifahanana M, Johansson SL, Cullen DM, Akhter MP et alChronic pancreatic inflammation induced by environmental tobacco smoke inhalation in rats. Am J Gastroenterol 2006; 101: 148–159.

    Article  CAS  PubMed  Google Scholar 

  9. Finnie IA, Campbell BJ, Taylor BA, Milton JD, Sadek SK, Yu LG et alStimulation of colonic mucin synthesis by corticosteroids and nicotine. Clin Sci 1996; 91: 359–364.

    Article  CAS  Google Scholar 

  10. Luk IS, Ho J, Wong WM, Yuen ST, Luk CT, Cho CH . Influence of chronic nicotine intake and acute ethanol challenge on gastric mucus level and blood flow in rabbits. Digestion 1994; 55: 399–404.

    Article  CAS  PubMed  Google Scholar 

  11. Chaturvedi P, Singh AP, Batra SK . Structure, evolution, and biology of the MUC4 mucin. FASEB J 2008; 22: 966–981.

    Article  CAS  PubMed  Google Scholar 

  12. Rachagani S, Torres MP, Moniaux N, Batra SK . Current status of mucins in the diagnosis and therapy of cancer. Biofactors 2009; 35: 509–527.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Kufe DW . Mucins in cancer: function, prognosis and therapy. Nat Rev Cancer 2009; 9: 874–885.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Bafna S, Kaur S, Batra SK . Membrane-bound mucins: the mechanistic basis for alterations in the growth and survival of cancer cells. Oncogene 2010; 29: 2893–2904.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Andrianifahanana M, Moniaux N, Schmied BM, Ringel J, Friess H, Hollingsworth MA et alMucin (MUC) gene expression in human pancreatic adenocarcinoma and chronic pancreatitis: a potential role of MUC4 as a tumor marker of diagnostic significance. Clin Cancer Res 2001; 7: 4033–4040.

    CAS  PubMed  Google Scholar 

  16. Swartz MJ, Batra SK, Varshney GC, Hollingsworth MA, Yeo CJ, Cameron JL et alMUC4 expression increases progressively in pancreatic intraepithelial neoplasia. Am J Clin Pathol 2002; 117: 791–796.

    Article  PubMed  Google Scholar 

  17. Chaturvedi P, Singh AP, Moniaux N, Senapati S, Chakraborty S, Meza JL et alMUC4 mucin potentiates pancreatic tumor cell proliferation, survival, and invasive properties and interferes with its interaction to extracellular matrix proteins. Mol Cancer Res 2007; 5: 309–320.

    Article  CAS  PubMed  Google Scholar 

  18. Moniaux N, Chaturvedi P, Varshney GC, Meza JL, Rodriguez-Sierra JF, Aubert JP et alHuman MUC4 mucin induces ultra-structural changes and tumorigenicity in pancreatic cancer cells. Br J Cancer 2007; 97: 345–357.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Singh AP, Moniaux N, Chauhan SC, Meza JL, Batra SK . Inhibition of MUC4 expression suppresses pancreatic tumor cell growth and metastasis. Cancer Res 2004; 64: 622–630.

    Article  CAS  PubMed  Google Scholar 

  20. Bafna S, Kaur S, Momi N, Batra SK . Pancreatic cancer cells resistance to gemcitabine: the role of MUC4 mucin. Br J Cancer 2009; 101: 1155–1161.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Mimeault M, Johansson SL, Senapati S, Momi N, Chakraborty S, Batra SK . MUC4 down-regulation reverses chemoresistance of pancreatic cancer stem/progenitor cells and their progenies. Cancer Lett 2010; 295: 69–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Lowenfels AB, Maisonneuve P . Risk factors for pancreatic cancer. J Cell Biochem 2005; 95: 649–656.

    Article  CAS  PubMed  Google Scholar 

  23. Choudhury A, Singh RK, Moniaux N, El-Metwally TH, Aubert JP, Batra SK . Retinoic acid-dependent transforming growth factor-beta 2-mediated induction of MUC4 mucin expression in human pancreatic tumor cells follows retinoic acid receptor-alpha signaling pathway. J Biol Chem 2000; 275: 33929–33936.

    Article  CAS  PubMed  Google Scholar 

  24. Lindstrom J, Schoepfer R, Whiting P . Molecular studies of the neuronal nicotinic acetylcholine receptor family. Mol Neurobiol 1987; 1: 281–337.

    Article  CAS  PubMed  Google Scholar 

  25. Egleton RD, Brown KC, Dasgupta P . Nicotinic acetylcholine receptors in cancer: multiple roles in proliferation and inhibition of apoptosis. Trends Pharmacol Sci 2008; 29: 151–158.

    Article  CAS  PubMed  Google Scholar 

  26. Ng J, Cantrell D . STAT3 is a serine kinase target in T lymphocytes. Interleukin 2 and T cell antigen receptor signals converge upon serine 727. J Biol Chem 1997; 272: 24542–24549.

    Article  CAS  PubMed  Google Scholar 

  27. Wen Z, Zhong Z, Darnell JE . Maximal activation of transcription by Stat1 and Stat3 requires both tyrosine and serine phosphorylation. Cell 1995; 82: 241–250.

    Article  CAS  PubMed  Google Scholar 

  28. Arredondo J, Chernyavsky AI, Jolkovsky DL, Pinkerton KE, Grando SA . Receptor-mediated tobacco toxicity: cooperation of the Ras/Raf-1/MEK1/ERK and JAK-2/STAT-3 pathways downstream of alpha7 nicotinic receptor in oral keratinocytes. FASEB J 2006; 20: 2093–2101.

    Article  CAS  PubMed  Google Scholar 

  29. Chaturvedi P, Singh AP, Chakraborty S, Chauhan SC, Bafna S, Meza JL et alMUC4 mucin interacts with and stabilizes the HER2 oncoprotein in human pancreatic cancer cells. Cancer Res 2008; 68: 2065–2070.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Provenzano PP, Keely PJ . The role of focal adhesion kinase in tumor initiation and progression. Cell Adh Migr 2009; 3: 347–350.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Wong HP, Li ZJ, Shin VY, Tai EK, Wu WK, Yu L et alEffects of cigarette smoking and restraint stress on human colon tumor growth in mice. Digestion 2009; 80: 209–214.

    Article  CAS  PubMed  Google Scholar 

  32. Majumdar AP, Davis GA, Dubick MA, Geokas MC . Nicotine stimulation of protein secretion from isolated rat pancreatic acini. Am J Physiol 1985; 248: G158–G163.

    CAS  PubMed  Google Scholar 

  33. Rubin H . Selective clonal expansion and microenvironmental permissiveness in tobacco carcinogenesis. Oncogene 2002; 21: 7392–7411.

    Article  CAS  PubMed  Google Scholar 

  34. Sheppard BJ, Williams M, Plummer HK, Schuller HM . Activation of voltage-operated Ca2+-channels in human small cell lung carcinoma by the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone. Int J Oncol 2000; 16: 513–518.

    CAS  PubMed  Google Scholar 

  35. Shen JX, Yakel JL . Nicotinic acetylcholine receptor-mediated calcium signaling in the nervous system. Acta Pharmacol Sin 2009; 30: 673–680.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Plummer HK, Dhar M, Schuller HM . Expression of the alpha7 nicotinic acetylcholine receptor in human lung cells. Respir Res 2005; 6: 29.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Dasgupta P, Rastogi S, Pillai S, Ordonez-Ercan D, Morris M, Haura E et alNicotine induces cell proliferation by beta-arrestin-mediated activation of Src and Rb-Raf-1 pathways. J Clin Invest 2006; 116: 2208–2217.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Marrero MB, Bencherif M . Convergence of alpha 7 nicotinic acetylcholine receptor-activated pathways for anti-apoptosis and anti-inflammation: central role for JAK2 activation of STAT3 and NF-kappaB. Brain Res 2009; 1256: 1–7.

    Article  CAS  PubMed  Google Scholar 

  39. Macha MA, Matta A, Chauhan SS, Siu KW, Ralhan R . Guggulsterone (GS) inhibits smokeless tobacco and nicotine-induced NF-kappaB and STAT3 pathways in head and neck cancer cells. Carcinogenesis 2011; 32: 368–380.

    Article  CAS  PubMed  Google Scholar 

  40. Chen RJ, Ho YS, Guo HR, Wang YJ . Rapid activation of Stat3 and ERK1/2 by nicotine modulates cell proliferation in human bladder cancer cells. Toxicol Sci 2008; 104: 283–293.

    Article  CAS  PubMed  Google Scholar 

  41. Chen RJ, Ho YS, Guo HR, Wang YJ . Long-term nicotine exposure-induced chemoresistance is mediated by activation of Stat3 and downregulation of ERK1/2 via nAChR and beta-adrenoceptors in human bladder cancer cells. Toxicol Sci 2010; 115: 118–130.

    Article  CAS  PubMed  Google Scholar 

  42. Andoh A, Shioya M, Nishida A, Bamba S, Tsujikawa T, Kim-Mitsuyama S et alExpression of IL-24, an activator of the JAK1/STAT3/SOCS3 cascade, is enhanced in inflammatory bowel disease. J Immunol 2009; 183: 687–695.

    Article  CAS  PubMed  Google Scholar 

  43. Mejias-Luque R, Peiro S, Vincent A, Van S I, de BC . IL-6 induces MUC4 expression through gp130/STAT3 pathway in gastric cancer cell lines. Biochim Biophys Acta 2008; 1783: 1728–1736.

    Article  CAS  PubMed  Google Scholar 

  44. Mejias-Luque R, Linden SK, Garrido M, Tye H, Najdovska M, Jenkins BJ et alInflammation modulates the expression of the intestinal mucins MUC2 and MUC4 in gastric tumors. Oncogene 2010; 29: 1753–1762.

    Article  CAS  PubMed  Google Scholar 

  45. Arredondo J, Chernyavsky AI, Marubio LM, Beaudet AL, Jolkovsky DL, Pinkerton KE et alReceptor-mediated tobacco toxicity: regulation of gene expression through alpha3beta2 nicotinic receptor in oral epithelial cells. Am J Pathol 2005; 166: 597–613.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Arredondo J, Chernyavsky AI, Grando SA . Nicotinic receptors mediate tumorigenic action of tobacco-derived nitrosamines on immortalized oral epithelial cells. Cancer Biol Ther 2006; 5: 511–517.

    Article  CAS  PubMed  Google Scholar 

  47. Andrianifahanana M, Agrawal A, Singh AP, Moniaux N, Van S I, Aubert JP et alSynergistic induction of the MUC4 mucin gene by interferon-gamma and retinoic acid in human pancreatic tumour cells involves a reprogramming of signalling pathways. Oncogene 2005; 24: 6143–6154.

    Article  CAS  PubMed  Google Scholar 

  48. Andrianifahanana M, Singh AP, Nemos C, Ponnusamy MP, Moniaux N, Mehta PP et alIFN-gamma-induced expression of MUC4 in pancreatic cancer cells is mediated by STAT-1 upregulation: a novel mechanism for IFN-gamma response. Oncogene 2007; 26: 7251–7261.

    Article  CAS  PubMed  Google Scholar 

  49. Ponnusamy MP, Singh AP, Jain M, Chakraborty S, Moniaux N, Batra SK . MUC4 activates HER2 signalling and enhances the motility of human ovarian cancer cells. Br J Cancer 2008; 99: 520–526.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Avizienyte E, Frame MC . Src and FAK signalling controls adhesion fate and the epithelial-to-mesenchymal transition. Curr Opin Cell Biol 2005; 17: 542–547.

    Article  CAS  PubMed  Google Scholar 

  51. Vadlamudi RK, Sahin AA, Adam L, Wang RA, Kumar R . Heregulin and HER2 signaling selectively activates c-Src phosphorylation at tyrosine 215. FEBS Lett 2003; 543: 76–80.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Ms Kristi L Berger for editing the manuscript. We acknowledge the invaluable technical support from Mr Erik Moore and Mrs Kavita Mallya. We also thank Janice A Tayor and James R Talaska, the members of the confocal laser scanning microscope core facility at UNMC, for their support. This work was, in part, supported by grants from the National Institutes of Health (RO1 CA133774,RO1 CA131944, RO1 CA78590, UO1 CA111294, P50 CA127297 and U54 CA163120).

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  1. N Momi and M P Ponnusamy: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE, USA

    N Momi, M P Ponnusamy, S Kaur, S Rachagani, M M Ouellette & S K Batra

  2. H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA

    S S Kunigal & S Chellappan

  3. Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE, USA

    M M Ouellette & S K Batra

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Correspondence to S K Batra.

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Momi, N., Ponnusamy, M., Kaur, S. et al. Nicotine/cigarette smoke promotes metastasis of pancreatic cancer through α7nAChR-mediated MUC4 upregulation. Oncogene 32, 1384–1395 (2013). https://doi.org/10.1038/onc.2012.163

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