Date of Award

Spring 4-24-2023

Document Type

Honors Project

Degree Name

Bachelor of Science

Department

Chemistry and Physics

Department Chair or Program Director

Dr. Janet Asper

First Advisor

Dr. Randall Reif

Major or Concentration

Biochemistry

Abstract

Apoptosis, commonly known as programmed cell death, constantly occurs in humans. As a cancer cell increases in acidity, apoptosis is induced. In healthy cells, proton pump proteins allow for H+ ions to permeate cellular membranes, regulating pH. However, proton pump inhibitors (PPIs), such as omeprazole, prevent proton movement. In previous studies, omeprazole induced cell death in Jurkat T lymphocytes; however, there was no confirmation of whether the cells died through apoptosis, or through necrosis, where the cell bursts. By using Annexin-V staining, the effects of omeprazole, dexlansoprazole, and esomeprazole on apoptosis induction can be measured. Cell death was observed by staining cells with propidium iodide (PI) dye and Annexin V-FITC proteins. In order to measure the extent of cell viability, cytosolic esterase activity was measured by staining cells with calcein-acetoxymethyl (AM) dye. Jurkat cells were exposed to PPIs omeprazole, dexlansoprazole, and esomeprazole for six hours and monitored for 30 hours to measure viability. Doxorubicin, a known chemotherapeutic, was also used as a positive control when testing apoptosis induction and viability. When imaged using fluorescent microscopy, any cell that was apoptotic fluoresced green and necrotic cells fluoresced red. With calcein-AM, if cells fluoresced, they were deemed viable, while nonfluorescent cells were deemed necrotic. At the 30-hour mark, dexlansoprazole showed the least viability, followed by doxorubicin, esomeprazole, and omeprazole, in comparison to the negative control. The low viability with dexlansoprazole indicated the need for a toxicity study using the same PPIs and exposure methods, to determine the optimal drug concentration.

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