Document Type

Thesis

Date of Award

8-2021

School/College

College of Science, Engineering, and Technology (COSET)

Degree Name

MS in Biology

Committee Chairperson

Desirée DJ Jackson

Committee Member 1

Shodimu-Emmanuel SEO Olufemi

Committee Member 2

Sonya SG Good, Hector HM Miranda

Keywords

Acrylamide, BAX, BCL-XL, BEAS-2B cells, PI3K/AKT pathway, Roxarsone

Abstract

Natural environmental contaminants and xenobiotic compounds consumed by humans are likely to be deleterious to human health. This study evaluates the effects of the organoarsenic compound roxarsone and the organic compound acrylamide on BEAS-2B cells. Arsenic occurs naturally in the environment in inorganic and organic forms. Inorganic arsenics are carcinogenic to humans. Organic arsenics (organoarsenics) are less toxic, yet their level of toxicity cannot be ignored. Roxarsone (4-hydroxy-3-nitrobenzenearsonate) is a compound used as a poultry feed additive and antimicrobial agent. It promotes cell proliferation and possesses angiogenesis properties. Roxarsone (ROX) can be found in chicken breast meat; along with a metabolite of ROX, thiolated roxarsone, which is more toxic than Roxarsone. Acrylamide is used in the manufacturing of paper, dye, and industrial products. It is also found in starchy food cooked at high temperatures. The body can be simultaneously exposed to ROX and acrylamide through food consumption. Acrylamide is known to cause cell damage, impair cell viability, and it affects cell proliferation. Experimental findings showed that 3.5 mM acrylamide destroys about half the population of BEAS-2B cells. Since acrylamide has deleterious effects on cells, it may be possible for ROX which stimulates cell proliferation, to ameliorate the deleterious effects of acrylamide. Previous studies showed that 10 µM Roxarsone induces cell proliferation. The specific aim of this study to demonstrate that the stimulation of BEAS-2B cell proliferation by 10 µM Roxarsone is not sufficient to protect the cells from damage induced by acrylamide. The first aim of this study is to demonstrate that treatment of BEAS-2B cells with ROX followed by treatment with acrylamide will alter cell morphology. The working hypothesis is that treatment of BEAS-2B cells with 10 µM ROX followed by 3.5 mM acrylamide will cause crenation, causing changes in cell morphology. The second aim of this study is to demonstrate that changes in cell morphology due to crenation will alter the PI3K/AKT pathway. The working hypothesis is that change in cell morphology will alter the proper functioning of genetic material and cellular responses, which is likely to destroy the PI3K/AKT cell survival pathway. Observation of cell morphology in response to ROX show that ROX induced cell proliferation in BEAS-2B cells with no morphological changes observed. Treatment of BEAS-2B cells with 10 µM ROX followed by 3.5 mM acrylamide changed cell morphology, which suggests that acrylamide is responsible for changes in cell morphology. Acrylamide-induced cell shrinkage and increased cell death were observed in treated BEAS-2B cells. ROX induced cell proliferation and caused significant upregulation of Ak strain transforming (AKT) levels after 24 hours of exposure. Cell proliferation stopped when the cells reached their maximum confluency at 48 hours. AKT and downstream genes in the PI3K/AKT pathway were analyzed in BEAS-2B cells treated with Roxarsone, or a combination of Roxarsone and acrylamide.

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