PARP-1 to the Rescue: A Biphasic Mechanism of UVB-Induced DNA Damage and Repair in Cultured Human Lens Epithelial Cells
Honors College Senior Thesis
It has been shown that UVB light damages the DNA of cells; however, it has yet to be shown how this occurs in lens epithelial cells (LECs) found on the anterior surface of the eye’s lens. The purpose of this research project was to explore UVB-induced DNA damage to LECs along with investigation of a related DNA repair complex. A DNA repair process for LECs is hypothesized to involve two components referred to as poly(ADP-ribose) polymerase-1 (PARP- 1) and poly(ADP-ribose) (PAR). The exact mechanisms for UVB induced DNA damage and its repair are not yet confirmed. To study these two events, cultured human LECs were exposed to UVB light and then incubated for various times after UVB exposure. Various assays were conducted to visualize the processes of DNA damage and repair. These assays included cell viability (MTT assay), the comet assay to detect single strand DNA breaks, reactive oxygen species (ROS) and superoxide anion indicators, as well as fluorescence immunocytochemistry with antibodies to PARP-1 and PAR and apoptosis detection. The goal was to discover how the lens DNA repair complex works in response to UVB induced cellular damage. The benefits of this project include a contribution to existing eye and cataract research.
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