Role of Versican in Epithelial Ovarian Carcinoma
thesisposted on 21.02.2013, 00:00 by Mark Desjardins
Epithelial ovarian carcinoma (EOC) is a leading cause of death from gynecologic malignancies, and the fifth leading cause of death in women. The extracellular matrix plays a key role in regulating cellular activities such as proliferation, adhesion, differentiation, and migration. Development of ovarian carcinoma can lead to the alteration of ECM components such as Versican (VCAN). Increased levels of versican have been found in primary epithelial ovarian tumors in pathology studies assessing the role of versican in EOC. The aim of this study was to elaborate the exact role that versican expression in migration and adhesion. Additionally, we sought to identify how versican expression affects multicellular aggregrates (spheroids) properties such as size, formation, and disaggregation. The importance of these multicellular aggregates has come to researchers attention as they have proven to be more resistant to traditional therapy than single cells. We have studied the expression of versican in cell lines expressing high levels of versican as well as lower levels of versican using QRT-PCR. The effect of versican on cellular migration was determined by performing the scratch wound healing assay, and monitoring wound closure over a 24 hour period. These results were corroborated using the modified boyden chamber method. Furthermore, silencing of versican expression in Skov-3, Dov-13, OVCA-429, and OVCA-433 cell lines using siRNA and shRNA resulted in an overall decrease in both single cell adhesion as well as multicellular adhesion on ECM components and on the LP-3 mesothelial monolayer. Additionally, the down-regulation of versican expression in Skov-3 and Dov-13 cell lines significantly inhibited their ability to disaggregate on the mesothelial monolayer and ECM components. We hypothesize that taken all together this data indicates that versican modulates EOC metastasis by increasing cellular adhesion, migration, spheroid formation, and multicellular disaggregation.