Skip to main content
SpringerLink
Log in

Role of nitric oxide in the induction of apoptosis by smokeless tobacco extract

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Smokeless tobacco usage is, a growing public health concern in the United States. Lesions of the oral cavity have been clearly linked to smokeless tobacco use. The objective of this study was to determine the biochemical effects of smokeless tobacco extract (STE) exposure upon hamster cheek pouch cell (HCPC-1) cultures. HCPC-1 cells were exposed to a 5 -fold dose-range of STE (0.5, 1.0 and 2.5%) over a time-course of 24-96 h. Following each exposure we measured various biochemical parameters of cell proliferation and cell death. Cell viability, cell cycle progression and S-phase DNA synthesis were measured as markers of cell proliferation. We measured lactate dehydrogenase leakage as a marker of cell membrane damage and cell death due to necrosis. No significant alterations were observed in cell cycle progression and cell proliferation as a result of exposure to STE. LDH measured colorimetrically indicated no significant effect with the lower doses (0.5, 1.0 and 2.5% STE). Apoptosis measured as the A0 peak and by the TUNEL procedure revealed that STE caused significant rates of apoptosis. Maximal apoptosis was noted between 48-96 h. In order to probe the mechanism further we measured the levels of nitrites as an indicator of nitric oxide (NO) in the media. NO levels were significantly elevated at the doses that caused an induction of apoptosis. The results from this study indicate that STE causes a dose-dependent induction of apoptosis and that this is mediated by nitric oxide.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Brownson RC, DiLorenzo TM, Van TM, Finger WW: Patterns of cigarette and smokeless tobacco use among children and adolescents. Prev Med 19: 170, 1990

    PubMed  Google Scholar 

  2. Tobacco use among high school students – United States. CDC, Atlanta, GA, 1990.

  3. Connolly GN, Winn DM, Hecht SS, Henningfield JE, Walker B, Hoffmann D: The reemergence of smokeless tobacco. N Engl J Med 314: 1020, 1986

    PubMed  Google Scholar 

  4. Giovino GA, Schooley MW, Zhu BP, Chrismon JH, Tomar SL, Peddicord JP, Merritt RK, Husten CG, Eriksen MP: Surveillance for selected tobacco-use behaviors – United States, 1900–1994. MMWR, CDC Surveill Summ 43: 1, 1994

    Google Scholar 

  5. Nelson DE, Tomar SL, Mowery P, Siegel PZ: Trends in smokeless tobacco use among men in four states, 1988 through 1993. Am J Publ Health 86: 1300, 1996

    Google Scholar 

  6. Kessler DA, Barnett PS, Witt A, Zeller MR, Mande JR, Schultz WB: The legal and scientific basis for FDA's assertion of jurisdiction over cigarettes and smokeless tobacco. JAMA 277: 405, 1997

    PubMed  Google Scholar 

  7. Report to Congress. Pursuant to the Comprehensive Smokeless Tobacco Health Education Act of 1986. Federal Trade Commission. Washington, DC, 1995

  8. Hoffmann D, Adams JD, Lisk D, Fisenne I, Brunnemann KD: Toxic and carcinogenic agents in dry and moist snuff. J Natl Cancer Inst 79: 1281, 1987

    PubMed  Google Scholar 

  9. Hoffmann D, Rivenson A, Chung F-L, Hecht SS: Nicotine-derived N-nitrosamines (TSNA) and their relevance in tobacco carcinogenesis. CRC Crit Rev Toxicol 21: 305, 1991

    Google Scholar 

  10. Kerr JF, Winterford CM, Harmon BV: Apoptosis. Its significance in cancer and cancer therapy [published erratum appears in Cancer 1994 Jun 15; 73(12: 3108]. Cancer 73: 2013

  11. Morris RG, Hargreaves AD, Duvall E, Wyllie AH: Hormoneinduced cell death. 2. Surface changes in thymocytes undergoing apoptosis. Am J Pathol 115: 426

  12. Gerschenson LE, Rotello RJ: Apoptosis: A different type of cell death. FASEB J 6: 2450, 1992

    PubMed  Google Scholar 

  13. Jacobson MD, Weil M, Raff MC: Programmed cell death in animal development. Cell 88: 347, 1997

    Article  PubMed  Google Scholar 

  14. Walker NI, Harmon BV, Gobe GC, Kerr JF: Patterns of cell death. Meth Ach Exp Pathol 13: 18, 1988

    Google Scholar 

  15. Williams GT: Programmed cell death: Apoptosis and oncogenesis. Cell 65: 1097, 1991

    PubMed  Google Scholar 

  16. Bursch W, Oberhammer F, Schulte HR: Cell death by apoptosis and its protective role against disease. Trends Pharmacol Sci 13: 245, 1992

    PubMed  Google Scholar 

  17. Patel T, Gores GJ: Apoptosis in liver transplantation: A mechanism contributing to immune modulation, preservation injury, neoplasia, and viral disease. Liver Transpl Surg 4: 42, 1998

    PubMed  Google Scholar 

  18. Sakagami H, Kuribayashi N, Iidi M, Sakagami T, Takeda M, Fukuchi K, Gomi K, Ohata H, Momose K, Kawazoe Y et al.: Induction of DNA fragmentation by tannin-and lignin-related substances. Anticancer Res 15: 2121, 1995

    PubMed  Google Scholar 

  19. Albina JE, Martin BA, Henry-WL J, Louis CA, Reichner JS: B cell lymphoma-2 transfected P815 cells resist reactive nitrogen intermediate-mediated macrophage-dependent cytotoxicity. J Immunol 157: 279, 1996.

    PubMed  Google Scholar 

  20. Albina JE, Reichner JS: Role of nitric oxide in mediation of macrophage cytotoxicity and apoptosis. Cancer Metastasis Rev 17: 39, 1998

    PubMed  Google Scholar 

  21. Messmer UK, Brune B: Nitric oxide-induced apoptosis: p53-dependent and p53-independent signalling, pathways. Biochem J 319: 299, 1996

    PubMed  Google Scholar 

  22. Brune B, Gotz C, Messmer UK, Sandau K, Hirvonen MR, Lapetina EG: Superoxide formation and macrophage resistance to nitric oxide-mediated apoptosis. J Biol Chem 272: 7253, 1997

    PubMed  Google Scholar 

  23. Oh JS, Cherrick HM, Park NH: Effect of snuff extract on the replication and synthesis of viral DNA and proteins in cells infected with herpes simplex virus. J Oral Maxillofac Surg 48: 373, 1990

    PubMed  Google Scholar 

  24. Hoffmann D, Adams JD, Brunnemann KD, Rivenson A, Hecht SS: Tobacco specific N-nitrosamines: Occurrence and bioassays. IARC Sci Publ 309, 1982

  25. Hoffmann D, Adams JD: Carcinogenic tobacco-specific Nnitrosamines in snuff and in the saliva of snuff dippers. Cancer Res 41: 4305, 1981

    PubMed  Google Scholar 

  26. Salvemini D, Seibert K, Masferrer JL, Settle SL, Misko TP, Currie MG, Needleman P: Nitric oxide and the cyclooxygenease pathway. Adv Prostagland Trombox Leukot Res 23: 491, 1995

    Google Scholar 

  27. Salvemini D, Misko TP, Masferrer IL, Seffiert K, Currie MG, Needleman P: Nitric oxide activates cyclooxygenase enzymes. Proc Natl Acad Sci USA 90: 7240, 1993

    PubMed  Google Scholar 

  28. Salvemini D: Regulation of cyclooxygenase enzymes by nitric oxide. Cell Mol Life Sci 53: 576, 1997

    PubMed  Google Scholar 

  29. Hortelano S, Dallaporta B, Zamami N, Hirsch T, Susin SA, Marzo I, Bosca L, Kroemer G: Nitric oxide induces apoptosis via triggering mitochondrial permeability transition. FEBS Lett 410: 373, 1997

    PubMed  Google Scholar 

  30. Xie K, Wang Y, Huang S, Xu L, Bielenberg D, Salas T, McConkey DJ, Jiang W, Fidler IJ: Nitric oxide-mediated apoptosis of K-1735 melanoma cells is associated with down regulation of Bcl-2. Oncogene 15: 771, 1997

    PubMed  Google Scholar 

  31. Blank KR, Rudoltz MS, Kao GD, Muschel RJ, McKenna WG: The molecular regulation of apoptosis and implications for radiation oncology. Int J Rad Biol 71: 455, 1997

    PubMed  Google Scholar 

  32. Blandino G, Strano S: BCL-2: The pendulum of the cell fate. J Exp Clin Cancer Res 16: 3, 1997

    PubMed  Google Scholar 

  33. Tang DG, Porter AT: Target to apoptosis: A hopeful weapon for prostate cancer. Prostate 32: 284, 1997

    PubMed  Google Scholar 

  34. Sakagami H, Satoh K, Fukuchi K, Kadofuku T, Gomi K, Nakamura K, Kuribayashi N, Sunaga S, Hirota N, Iida M, Makino Y, Kojima T, Shimura H, Takeda M: Effect of methionine depletion on growth and apoptosis in various tumor cell lines. Anticancer Res 17: 2407, 1997

    PubMed  Google Scholar 

  35. Maneckjee R, Minna JD: Opioids induce while nicotine suppresses apoptosis in human lung cancer cells. Cell Growth Diff 5: 1033, 1994

    PubMed  Google Scholar 

  36. Wright SC, Zhong J, Zheng H, Larrick JW: Nicotine inhibition of apoptosis suggests a role in tumor promotion. FASEB J 7: 1045, 1993

    PubMed  Google Scholar 

  37. Yan GM, Lin SZ, Irwin RP, Paul SM: Activation of muscarinic cholinergic receptors blocks apoptosis of cultured cerebellar granule neurons. Mol Pharmacol 47: 248, 1995

    PubMed  Google Scholar 

  38. Des RC, Dische S, Saunders MI: The problem of cigarette smoking in radiotherapy for cancer in the head and neck. Clin Oncol R Coll Radiol 4: 214, 1992

    PubMed  Google Scholar 

  39. Browman GP, Wong G, Hodson I, Sathya J, Russell R, McAlpine L, Skingley P, Levine MN: Influence of cigarette smoking on the efficacy of radiation therapy in head and neck cancer [see comments]. N Engl J Med 328, 159, 1993

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE, 68198, USA

    Raja S. Mangipudy & Jamboor K. Vishwanatha

Authors
  1. Raja S. Mangipudy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jamboor K. Vishwanatha
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mangipudy, R.S., Vishwanatha, J.K. Role of nitric oxide in the induction of apoptosis by smokeless tobacco extract. Mol Cell Biochem 200, 51–57 (1999). https://doi.org/10.1023/A:1006985700851

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006985700851

Search

Navigation