 | Eiffel John Q. Manzano University of Guam (Biology '09)Mentor(s)Lori A. White, Ph.D. Kyle A. Murphy Department of Biochemistry and Microbiology Rutgers University |
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| Development of zebrafish (Danio rerio) as a model for studying effects of ultraviolet (UVB) light exposure on melanoma progression | ||
| The A2058 human melanoma cell line has been used by our laboratory for studies of matrix metalloproteinase (MMP) expression and elucidation of the role of the aryl hydrocarbon receptor (AhR) pathway in tumorigenesis. MMPs are endopeptidases that degrade the protein components of the extracellular matrix and basement membrane. Expression of these enzymes is necessary for cellular migration and invasion. Recently, other laboratories have demonstrated that the zebrafish can be used as a model to examine melanoma migration and invasion. As our laboratory is interested in how environmental exposure contributes to melanoma metastasis, we have decided to use the zebrafish model to examine the effect of these exposures in a whole animal level. The goal of this project is to gather preliminary data examining the migration and invasion of A2058 cells in zebrafish. A protocol has been generated to label these cells with CM-DiI, a fluorescent dye that would track the distribution of the cells in the zebrafish embryos. Briefly, post-labeling A2058 cells that were replated have been found to retain their fluorescence and viability more effectively than those suspended in media. We have generated preliminary data demonstrating the effects of ultraviolet (UVB) light on CYP1A1, a protein significant for drug metabolism, and MMP expression in the zebrafish. In addition, an appropriate UVB exposure for the zebrafish has been identified for use in later studies. To demonstrate changes in MMP expression, total RNA was isolated from the embryos, and was used as template for quantitative real time RT-PCR. These results would serve as a positive control when the process of injecting the A2058 cells in zebrafish is performed in the future. |