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The Effect of TGF-beta Stimulation on Retinal Pigment Epithelium Cell Transformation

CSEF · 2008 Biochemistry/ Molecular Biology

Oscar Calzada · Bravo (Francisco) Medical Magnet High School, Los Angeles, Los Angeles County

Overview

Objectives/Goals Retinal pigment epithelium (RPE) cells are located outside the neuro-sensory retina that nourishes photoreceptor cells in the human eye. Transformed RPE plays an important role in the pathogenesis of proliferative vitreoretinopathy (PVR). Smooth-Muscle Actin (SMA) is a critical marker for RPE transdifferentiation and we hope to elucidate the factors that cause and are involved in RPE cell transformation. Methods/Materials A cell culture of retinal pigment epithelium cells from human fetal eye is grown continuously in the lab. After pre-treating RPE cells with Transforming Growth Factor-beta (TGF-beta) and 5AZA (DNA methylation inhibitor), we performed RNA isolation, followed by reverse transcription, Real-time Polymerase Chain Reaction (Real-time PCR), using SMA specific primer. Results Compared to our control, TGF-beta pre-treated cells see a fold-increase in RNA levels of SMA, whereas 5AZA pre-treated cells see a fold-decrease in the expression of SMA mRNA. Conclusions/Discussion Stimulating RPE cells with TGF-beta can increase SMA RNA expression, while treatment of RPE cells with 5AZA, inhibits SMA mRNA expression.

Summary statement

To determine the role played by TGF-beta protein in retinal pigment epithelium (RPE) cell transformation and its regulation by 5-AZA.

Help received

Used lab equipment at the Doheny Vision and Research Center of the University of Southern California under the supervision of Dr. Shikun He; Participant in the Science Technology and Research (STAR II) internship program.

Competition history

  • CSEF 2008 Biochemistry/ Molecular Biology · Entry S0404

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