Kovach, Margaret J.
Potts, Gretchen E.; Richards, Sean
University of Tennessee at Chattanooga
Place of Publication
The effects of smoking have long been studied; however, with the rise of electronic cigarettes there is a growing interest in the effects of this smoking substitute. Marketed as a safer alternative to tobacco cigarettes, the popularity of electronic cigarette (e-cigarette) use has dramatically increased. This has demanded regulatory methods to be put into effect, however, scientific study is only beginning on the new fad to determine the impacts of its use on the human body. The present study aims to examine the biological effects of lung cell exposure to tobacco alkaloids found in electronic cigarette filling solutions. We hypothesize that e-cigarette alkaloids, namely nicotine, cotinine, myosmine, and anabasine singularly and in combination can contribute to changes in normal cell proliferation. To address the aims of this study, two directions for evaluating cell proliferation by luminescent output were pursued. One of these was a genetically engineered stable lung cell line with the pCMVlux system, whose plasmid confers an auto-bioluminescent phenotype, allowing for analysis of cell proliferation through detection of a luminescent signal. The other direction of pursuit was the Promega Cell-Titer Glo® Luminescent Viability Assay. Experimental conditions were evaluated in triplicate after 48 and 96 hours of exposure. Potential synergistic and antagonistic effects were examined by alkaloid exposure in combination. A two-tailed, 2-sample t-test (α = 0.05) was used to evaluate statistically significant differences between experimental conditions and controls. Statistically significant differences in cell proliferation were found in exposures to all four alkaloids, but trends of their effects in combination were difficult to discriminate. Anabasine and myosmine have potential synergistic effects in stimulating cell growth, while cotinine has the potential to act as an antagonist against their activity increasing cellular proliferation.
I would like to thank Dr. Margaret Kovach, who graciously sacrificed time and effort to serve not only as my Departmental Honors project director, but also as my mentor through all my time as a student in her lab. I would like to also thank Dr. Sean Richards and Dr. Gretchen Potts for kindly serving on my committee. I would also like to thank Dr. Potts, specifically, for providing the alkaloids for this continuing study. This research was supported in part by the URaCE SEARCH Award.
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.
Alkaloids; Cell proliferation; Electronic cigarettes
Morrison, Olivia, "Development of an auto-bioluminescent lung cell line for evaluation of in vitro effects of e-cigarette compounds on normal cell proliferation" (2020). Honors Theses.