Project Director

Kovach, Margaret J.

Department Examiner

Carver, Ethan A.; Potts, Gretchen E.

Department

Dept. of Biological and Environmental Sciences

Publisher

University of Tennessee at Chattanooga

Place of Publication

Chattanooga (Tenn.)

Abstract

Marketed as a safer alternative to tobacco products, the popularity of e-cigarette use is on the rise. However, little is known about the potential health risks associated with their use. Many e-cigarette filling solutions are known to contain significant levels of tobacco alkaloids, including nicotine, anabasine, myosmine and cotinine. Using a panel of lung cell cultures distinguished by differences in sex and disease status, this study addresses the in vitro effects of common tobacco alkaloids found in e-cigarettes on cell proliferation and gene expression. We hypothesize that alkaloid exposure of lung cells is associated with abnormal proliferation and gene expression, and predict that cellular response to the alkaloids will present in a sex-specific manner. Alkaloid exposure on each lung cell line was evaluated at 1 µg/mL, 10 µg/mL, and 100 µg/mL concentrations throughout a 10-day time course. Cellular proliferation was measured daily using the CellTiter-Glo® Luminescent Viability Assay, and RNA isolated after 48 and 96-hour time points for gene expression analysis of 10 cancer biomarkers by qRT-PCR. Findings indicate anabasine and myosmine display a significant (p<0.05) inhibitory effect on cellular proliferation and gene expression, whereas the effect of cotinine and nicotine was minimal. Each cell line had an inherent, characteristic pattern of cellular proliferation in response to alkaloid exposure. Notably, the cancer cell line demonstrated more variability among replicates, which we attribute to the non-clonal nature of this cell line, and the female cell line displayed increased susceptibility to toxicity by the higher alkaloid concentrations. Significant differences in gene expression (p<0.05) were noted for AHR, CEACAM6, CYP1A1, MDM2, TP53, ALDH3A1 and GPX2. Correlation of abnormal patterns of cell proliferation and differential gene expression with risk for disease are discussed.

Acknowledgments

I would like to thank Dr. Margaret Kovach, who kindly sacrificed time and effort to serve not only as my Departmental Honors project director, but also as my mentor. I would like to also thank Dr. Ethan Carver and Dr. Gretchen Potts for kindly serving on my committee, as well as for Dr. Potts, specifically, for providing the alkaloids for this study. This research was supported in part by the UTC Provost Student Research Award and the UTC CRISP Award

Degree

B. S.; An honors thesis submitted to the faculty of the University of Tennessee at Chattanooga in partial fulfillment of the requirements of the degree of Bachelor of Science.

Date

8-2017

Subject

Electronic cigarettes -- Analysis; Electronic cigarettes -- Health aspects

Keyword

Tobacco; Alkaloids; E-cigarette; Gene expression; lung cell culture; Gender

Discipline

Environmental Sciences

Document Type

Theses

Extent

iii, 100 leaves

Language

English

Rights

Under copyright.

License

http://creativecommons.org/licenses/by-nc-nd/3.0/

Date Available

8-5-2017

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