Date of Award

2001

Document Type

Thesis

Degree Name

Master of Science

Department

Biology

First Advisor

Valerie M. Kish

Abstract

Overexpression of matrix metalloproteinase 2, MMP-2, is correlated with highgrade gliomas in vivo. The purpose of this study was to investigate whether MMP-2 overexpression in glioma cell lines leads to increased invasion of glioma cells in vitro. The entire MMP-2 gene was isolated from U87, a glioma cell line known to have high expression of wild-type MMP-2. This :tvIMP-2 gene was inserted in a sense and antisense orientation into pCR3 .1 vectors with a CMV promoter to ensure high levels of expression. Glioma cell lines were transfected with the sense and antisense constructs, as well as the pCR3.l vector alone to serve as a control. Zymography and Western blot analysis of conditioned media taken from glioma cell lines U251, T98, and LNZ-308 transfected with sense MMP-2 revealed an overproduction ofMMP-2 relative to controls. The invasiveness of these transfected cell lines was evaluated using an in vitro invasion assay. These assays revealed significantly more invasion by cells that were overproducing MMP-2 relative to cells containing either vector alone (control) or vector containing the antisense construct. These results support the hypothesis that glioma cell lines overexpressing MMP2 are more invasive in vitro.

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