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Transformation of Non-Cancerous Human Breast Epithelial Cell Line MCF10A by the Tobacco-Specific Carcinogen NNK

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Abstract

Repeated treatments of non-cancerous human breast epithelial cells MCF10A with a low dose of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) induced the development of cancerous cells. NNK-transformed MCF10A cells acquired cancerous properties including anchorage-independent cell growth and increased cell motility. Cellular transformation of MCF10A cells was accompanied by a loss of responsiveness to 17β-estradiol and decreased rate of cell proliferation. NNK-transformed MCF10A cells were also tumorigenic in immunodifficient mice. Studies of changes in the regulation of intracellular signaling pathways revealed that the upstream Erk pathway was down-regulated in the NNK-transformed cells. Our data provide the first evidence suggesting that the tobacco carcinogen NNK is competent to induce malignant transformation of non-cancerous human breast epithelial cells. Our findings suggest that the tobacco carcinogen NNK may contribute to early events in human breast carcinogenesis.

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References

  1. Hecht SS: Tobacco smoke carcinogens and lung cancer. J Natl Cancer Inst 91: 1194–1210, 1999

    Google Scholar 

  2. Hecht SS: Recent studies on mechanisms of bioactivation and detoxification of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a tobacco, specific lung carcinogen. Crit Rev Toxicol 26: 163–181, 1996

    Google Scholar 

  3. Jee SH, Ohrr H, Kim IS: Effects of husbands' smoking on the incidence of lung cancer in Korean women. Int J Epidemiol 28: 824–828, 1999

    Google Scholar 

  4. Lash TL, Aschengrau A: Active and passive cigarette smoking and the occurrence of breast cancer. Am J Epidemiol 149: 5–12, 1999

    Google Scholar 

  5. Morabia A: Smoking (active and passive) and breast cancer: epidemiologic evidence up to June 2001. Env Mol Mutagenesis 39: 89–95, 2002

    Google Scholar 

  6. Hecht SS: Tobacco smoke carcinogens and breast cancer. Env Mol Mutagenesis 39: 119–126, 2002

    Google Scholar 

  7. Perera FP, Estabrook A, Hewer A, Channing K, Rundle A, Mooney LA, Whyatt R, Phillips DH: Carcinogen, DNA adducts in human breast tissue. Cancer Epidemiol Biomarkers Prev 4: 233–238, 1995

    Google Scholar 

  8. Wu Z, Upadhyaya P, Carmella SG, Hecht SS, Zimmerman CL: Disposition of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in bile duct-cannulated rats: stereoselective metabolism and tissue distribution. Carcinogenesis 23: 171–179, 2002

    Google Scholar 

  9. Schuller HM, Orloff M: Tobacco-specific carcinogenic nitrosamines: ligands for nicotinic acetylcholine receptors in human lung cancer cells. Biochem Pharmacol 55: 1377–1384, 1998

    Google Scholar 

  10. Schuller HM, Tithof PK, Williams M, Plummer III H: The tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone is a beta-adrenergic agonist and stimulates DNA synthesis in lung adenocarcinoma via beta adrenergicreceptor-mediated release of arachidonic acid. Cancer Res 15(59): 4510–4515, 1999

    Google Scholar 

  11. Schuller HM, Porter B, Riechert A: Beta-adrenergic modulation of NNK-induced lung carcinogenesis in hamsters. J Cancer Res Clin Oncol 126: 624–630, 2000

    Google Scholar 

  12. Schuller HM, Jull BA, Sheppard BJ, Plummer III H: Interaction of tobacco-specific toxicants with the neuronal alpha(7) nicotinic acetylcholine receptor and its associated mitogenic signal transduction pathway: potential role in lung carcinogenesis and pediatric lung disorders. Eur J Pharmacol 393: 265–277, 2000

    Google Scholar 

  13. Jull BA, Plummer III HK, Schuller HM: Nicotinic receptormediated activation by the tobacco-specific nitrosamine NNK of a Raf-1/MAP kinase pathway, resulting in phosphorylation of c-myc in human small cell lung carcinoma cells and pulmonary neuroendocrine cells. J Cancer Res Clin Oncol 127: 707–717, 2001

    Google Scholar 

  14. Sabourin CL, Wang QS, Ralston SL, Evans J, Coate J, Herzog CR, Jones SL, Weghorst CM, Kelloff GJ, Lubet RA, You M, Stoner GD: Expression of cell cycle proteins in 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanoneinduced mouse lung tumors. Exp Lung Res 24: 499–521, 1998

    Google Scholar 

  15. Soule HD, Maloney TM, Wolman SR, Peterson Jr WD, Brenz R, McGrath CM, Russo J, Pauley RJ, Jones RF, Brooks SC: Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10. Cancer Res 50: 6075–6086, 1990

    Google Scholar 

  16. Hecht SS, Trushin N, Reid-Quinn CA, Burak ES, Jones AB, Southers JL, Gombar CT, Carmella SG, Anderson LM, Rice JM: Metabolism of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butane in the patas monkey; pharmacokinetics and characterization of glucuronide metabolites. Carcinogenesis 14: 229–236, 1993

    Google Scholar 

  17. Sommers CL, Papageorge A, Wilding G, Gelmann EP: Growth properties and tumorigenesis of MCF7 cells transfected with isogenic mutants of rash. Cancer Res 50: 67–71, 1990

    Google Scholar 

  18. O'Leary JJ, Mehta C, Hall DJ, Rosenberg A: Quantitation of [3H]-thymidine uptake by stimulated human lymphocytes. Cell Tissue Kinet 13: 21–32, 1980

  19. Song MK, Krutzsch H, Hankins WD, Richards WL, Thorgeirsson SS: Rapid determination of DNA synthesis in adherent cells grown in microtiter plates. Exp Cell Res 156: 271–276, 1985

    Google Scholar 

  20. Wang HCR, Fecteau KA: Detection of a novel quiescencedependent protein kinase. J Biol Chem 275: 25850–25857, 2000

    Google Scholar 

  21. Fecteau KA, Mei J, Wang HCR: Differential modulation of signaling pathways and apoptosis of ras-transformed 10T1/2 cells by the depsipeptide FR901228. J Pharmacol Exp Ther 300: 890–899, 2002

    Google Scholar 

  22. Kahn D, Andrieu JM, Dray F: Evaluation of some binding characteristics using dextran-coated charcoal to separate the bound and free fractions. Immunochemistry 11: 327–332, 1974

    Google Scholar 

  23. Schata F, Gurpide E: Effects of estradiol on prostaglandin F2? levels in primary monolayer cultures of epithelial cells from human proliferative endometrium. Endocrinol 113: 1274–1279, 1983

    Google Scholar 

  24. Jeffrey JJ, Ehlich LS, Roswit WT: Serotonin: an inducer of collagenase in myometrial smooth muscle cells. J Cell Physiol 146: 399–406, 1991

    Google Scholar 

  25. Repesh LA: A new in vitro assay for quantitating tumor cell invasion. Invasion Metastasis 9: 192–208, 1989

    Google Scholar 

  26. Sun XG, Rotenberg SA: Overexpression of protein kinase C? in MCF-10A human breast cells engenders dramatic alterations in morphology, proliferation, and motility. Cell Grow Diff 10: 343–352, 1999

    Google Scholar 

  27. Nollet F, Berx G, van Roy F: The role of the E-cadherin/ catenin adhesion complex in the development and progression of cancer. Mol Cell Biol Res Commun 2: 77–85, 1999

    Google Scholar 

  28. Critchley DR: Focal adhesions-the cytoskeletal connection. Curr Opin Cell Biol 12: 133–139, 2000

    Google Scholar 

  29. Campbell SL, Khosravi-Far R, Rossman KL, Clark GL, Der CJ: Increased complexity of Ras signaling. Oncogene 17: 1395–1413, 1998

    Google Scholar 

  30. Wang HCR, Erikson RL: Activation of protein serine/ threonine kinases p42, p63 and 87 in Rous sarcoma virustransformed cells: signal transduction/transformationdependent MBP kinases. Mol Biol Cell 3: 1329–1337, 1992

    Google Scholar 

  31. Mehta RR, Graves JM, Hart GD, Shilkaitis A, Das Gupta TK: Growth and metastasis of human breast carcinomas with Matrigel in athymic mice. Breast Cancer Res Treat 25: 65–71, 1993

    Google Scholar 

  32. Mullen P, Ritchie A, Langdon SP, Miller WR: Effect of Matrigel on the tumorigenicity of human breast and ovarian carcinoma cell lines. Int J Cancer 67: 816–820, 1996

    Google Scholar 

  33. Clark GJ, Kinch MS, Gilmer TM, Burridge K, Der CJ: Overexpression of the Ras-related TC21/R-Ras2 protein may contribute to the development of human breast cancers. Oncogene 12: 169–176, 1995

    Google Scholar 

  34. Freedman VH, Shin SI: Cellular tumorigenicity in nude mice: correlation with cell growth in semi-solid medium. Cell 3: 355–359, 1974

    Google Scholar 

  35. Basolo F, Elliott J, Tait L, Chen XQ, Maloney T, Russo IH, Pauley R, Momiki S, Caamano J, Klein-Szanto AJP, Koszalka M, Russo J: Transformation of human breast epithelial cells by c-Ha-ras oncogene. Mol Carcinog 4: 25–35, 1991

    Google Scholar 

  36. English J, Pearson G, Wilsbacher J, Swantek J, Karandikar M, Xu S, Cobb MH: New insights into the control of MAP kinase pathways. Exp Cell Res 253: 255–270, 1999

    Google Scholar 

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Authors and Affiliations

  1. Department of Comparative Medicine, The University of Tennessee, Knoxville, TN, USA

    Jianxun Mei, Hongbo Hu, Ping Song & Hwa-Chain R. Wang

  2. Department of Pathology, College of Veterinary Medicine, The University of Tennessee, Knoxville, TN, USA

    Michael McEntee & Howard Plummer III

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  1. Jianxun Mei
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  2. Hongbo Hu
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  3. Michael McEntee
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  4. Howard Plummer III
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  5. Ping Song
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  6. Hwa-Chain R. Wang
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Mei, J., Hu, H., McEntee, M. et al. Transformation of Non-Cancerous Human Breast Epithelial Cell Line MCF10A by the Tobacco-Specific Carcinogen NNK. Breast Cancer Res Treat 79, 95–105 (2003). https://doi.org/10.1023/A:1023326121951

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