Document Type

Thesis

Date of Award

5-2007

School/College

College of Science, Engineering, and Technology (COSET)

Degree Name

MS in Biology

First Advisor

G. T. Ramesh, Ph.D., Advisor

Abstract

Skin is one of the primary targets for exposure to uranium in the work place through contact in contaminated environments. It is for this reason that the following study is undertaken to investigate the toxic effects of uranyl acetate (UA) in human BJ Foreskin cells. The objective of this study is to analyze the signaling pathway resulting in oxidative stress induced inhibition of cell proliferation by uranyl acetate in BJ Foreskin. The hypothesis is that a dose of 1 mM of uranyl acetate will approximately decrease cell proliferation by 50% compared to control cells. Uranyl acetate is a water-soluble compound made from depleted uranium (DU). Previous studies have shown cancer mortality is significantly higher for people living in counties of Colorado where uranium tailing was used as a construction fill. More recently, a significant increase in gastric cancer was reported in counties of New Mexico with significant deposits of uranium or uranium tailings. This study revealed a significant increase in cancer of the prostrate, pancreas, stomach and colon. To date, there has been no report describing the toxicological effects of uranium on skin or epithelium. 1 2 The following experiments and assays were used to gather information helpful in determining the answer to the proposed hypothesis. Reactive Oxygen Species (ROS) is the most important class of metabolites that are unstable and potentially reactive. ROS will be measured by DiChloroFluorescein [5 -(and -6)- carboxy - 2,7 - dichloro - dihydroxy fluorescein diacetate (DCF), a fluorescent dye. The determination of cellular proliferation, viability and activation are key areas studied using the MTT (3-[4, 5-diMethylThiazol-2-ylJ-2, 5-diphenylTetrazolium bromide) assay. A live/dead cell kit was ran for distinguishing between cell growth and cell death. The kit used a Live-Dye, cell-permeable green fluorescent dye to stain live cells. Antioxidants were used as a countermeasure in order for the cell to be protected from outside exposures.

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