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Inhibition of nitric oxide-induced apoptosis by nicotine in oral epithelial cells

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Abstract

Development of oral cancer is clearly linked to the usage of smokeless tobacco. The molecular mechanisms involved in this process are however not well understood. Toward this goal, we investigated the effect of smokeless tobacco exposure on apoptosis of oral epithelial cells. Exposure of oral epithelial cells to smokeless tobacco extract (STE) induces apoptosis in a dose-dependent manner, until a threshold level of nicotine is achieved upon which apoptosis is inhibited. 1 mM of nicotine is able to inhibit apoptosis significantly induced by STE in these oral cells. Exposure of cells to nicotine alone has no effect on apoptosis, but nicotine inhibits apoptosis induced by other agents present in STE. In this study we show that, the anti-apoptotic action of nicotine is specifically associated with down-regulation of nitric oxide (NO) production. Using specific inducers of NO, we have demonstrated that inhibition of apoptosis by nicotine is through down-regulation of NO production. Further, we observed that nicotine clearly acts as a sink of NO radicals, shown using peroxynitrite generator (SIN-1) in conjunction or absence of radical scavengers. Nicotine thus causes most damage in transformed epithelial cells as depicted by accumulation of nitrotyrosine in a 3-NT ELISA assay. Inhibition of apoptosis is a hallmark in tumor progression and propels development of cancer. It may further result in functional loss of apoptotic effector mechanisms in the transformed cells. Thus, our data clearly indicates that inhibition of NO-induced apoptosis by nicotine may lead to tobacco-induced oral carcinogenesis, and implies careful development of modalities in tobacco cessation programs.

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Acknowledgments

Research described in this article was supported by a grant from the RMG, PM International to JKV. We are thankful to Drs. Wong DTW and Lekic PC for the gift of valuable cell lines for our research, and technical assistance by Ms. Judy Kanungo (AGB lab), Ms. Petrina Setiloane (AGB Lab), and Mr. Randall Davis (JKV lab).

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Authors and Affiliations

  1. Department of Oral Biology, Dental Diagnostic and Surgical Sciences, University of Manitoba, Health Science Center Campus, Winnipeg, MB, Canada, R3E0W2

    Abhijit G. Banerjee

  2. Karpagam Arts and Science College, Coimbatore, India

    Velliyur K. Gopalakrishnan

  3. Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, TX, 76107, USA

    Jamboor K. Vishwanatha

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  1. Abhijit G. Banerjee
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  2. Velliyur K. Gopalakrishnan
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  3. Jamboor K. Vishwanatha
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Correspondence to Abhijit G. Banerjee.

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Abhijit G. Banerjee and Velliyur K. Gopalakrishnan—contributed equally.

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Banerjee, A.G., Gopalakrishnan, V.K. & Vishwanatha, J.K. Inhibition of nitric oxide-induced apoptosis by nicotine in oral epithelial cells . Mol Cell Biochem 305, 113–121 (2007). https://doi.org/10.1007/s11010-007-9534-2

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