Date of Award

2016

Degree Type

Thesis

Degree Name

Master of Science in Biomedical Sciences

First Advisor

Ruth Borghaei, Ph.D. Thesis Advisor

Second Advisor

Farzaneh Daghigh, Ph.D.

Third Advisor

Dawn Shell, Ph.D.

Abstract

Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases with the unique ability to breakdown virtually the entire extracellular matrix (ECM). Through ECM remodeling, MMPs play an important role in normal development, tissue repair, angiogenesis, and apoptosis. Studies have shown that unregulated MMP expression plays a role in many cancers and chronic inflammatory diseases. Previous research has used gene sequence analysis ofthe MMP-1 promoter to identify a putative ZBP-89 binding site at -1969 bp. Chromatin immuneprecipitation experiments showed that both ZBP-89 and Re!A (p65) could bind to this binding site. In this thesis research, transfection experiments were used to explore the role of ZBP-89 and p65 in regulating MMP-1 gene expression. Two versions of a MMP-1 plasmid were used: a "Long MMP-1 plasmid" with a longer MMP-1 promoter (2.2 kb) that contains the distal putative binding site and a "Short MMP-1 plasmid with a shorter promoter (1.1 kb) that does not. Results showed that ZBP-89 alone can increase long MMP-1 plasmid expression in COS-1 cells and ZBP- 89 and p65 synergistically increase long MMP-1 plasmid expression in a dose-dependent manner. This suggests that ZBP-89 may cooperate with NFkB-p65 in MMP-1 gene regulation. ZBP-89 and p65 did not increase short MMP-1 plasmid expression in COS-1 cells or A549 cells. These results support our hypothesis that ZBP-89 and p65 work directly through the putative binding site at -1969 bp. This research further expands our understanding of MMP-1 gene regulation and can aid the development of MMP targeted therapy.

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