College of Arts & Sciences
mRNA expression of P-glycoprotein (Pgp) in MDCKII (MDR1 & wild type) and Caco-2 cell lines using TaqMan probe RT-PCR
De vika Ba xi COAS Biological Sciences, BS/MD
Dr . Jo se ph B entz Faculty Mentor Biology Ms. Guoying T ai Glaxosmithkline Ms. Neha Akella Glaxosmithkline
Poster Session B
Multidrug resistance is an ever-present concern in the world of modern medicine. Integral membrane transport proteins, such as P-glycoprotein (Pgp), play a major role in this phenomenon. This protein plays an essential role in drug efflux from the target site and bloodstream. To better understand Pgp, a relationship between amount of protein and function must be established. The first set of experiments was designed to determine the amount of intracellular mRNA that codes for Pgp. Three cell lines were used: Caco-2 and two Madin-Darby Canine Kidney Cell line derivatives. MDCKII MDR1 cells were transfected with human MDR1 gene to overexpress Pgp and MDCKII wild type cells served as a control. RNA was isolated from these cell lines and run through a TaqMan probe RTPCR with MDRI sequence specific probes, and the amount of mRNA for Pgp was determined for each cell line. A second set of Mass Spec analysis experiments will follow to quantify Pgp in the above cell lines. After further research, a relationship will be established between levels of Pgp and corresponding activity. By establishing this relationship, the kinetics of this transporter will be better understood and will help the research against multidrug resistance.
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