Date of Award

2009

Degree Type

Thesis

Degree Name

Master of Science

Program

Biochemistry

Supervisor

David Rodenhiser

Second Advisor

Alan Tuck

Third Advisor

Mellissa Mann

Abstract

Epithelial-mesenchymal transition (EMT) has been implicated in the process of

breast cancer metastasis, and ZEB2 has been implicated in EMT. Some evidence exists

suggesting that epigenetic DNA methylation modifications may be involved in EMT;

however, the specific contribution of DNA methylation to ZEB2 regulation is poorly

understood. I hypothesize that aberrant hypomethylation of the ZEB2 promoter results in

increasedexpressionofZEB2incellsthathavegonethroughEMT. Iobserveddecreased

ZEB2 promoter methylation and increased ZEB2 expression in vitro in mesenchymal-like /

breast cancer cells. Functional assays performed by treating cells with epigenetic inhibitors revealed ZEB2 demethylation that correlated with ZEB2 re-expression in a cell line sub-population. As well, a region flanking the ZEB2 gene was defined as having promoter activity, and this activity was sensitive to in vitro DNA methylation. These results support my hypothesis that DNA methylation may be involved in the regulation of ZEB2 expression.

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