WNT5A Modulates Integrins Expression Of Human Ovarian Cancer Cells

Metastasize of epithelail ovarian cancer (EOC) occurs by forming spheroids in peritoneum cavityaccompanied with adherence and disaggregation of cells to mesothelium. Both cell-cell adhesionand cell-ECM interacting molecules play a role in this process and a number of cell adhesionmolecules such as integrins have been suggested. WNT5A protein is an important regulator ofcancer cell behavior and progression in different types of cancer. Here we sought to determine themodulatory role of Wnt5A on integrin expression by human ovarian cancer cell line SKOV3 inmonolayer and spheroid model. Cells were stably transfected with pcDNA3.2/hWNT5A(Wnt5A/SKOV-3) construct or empty vector (mock). Wnt5A, N-cadherin, snail and fibronectinexpression levels were assessed by qPCR. Integrins expression was analyzed by using integrin arraykit. Effect of ECM components such as collagen type- I and IV, laminin and fibronectin on spheroidformation was assessed by mixing them with Mock and Wnt5A/SKOV-3 cells. Cell’s migration wasanalyzed by using both scratch and transwell assay. Moreover, BOX-5 as a Wnt5A antagonist wasused to assess the effect of Wnt5A blockage on integrin expression and migration of Wnt5A/SKOV-3cells. Wnt5A overexpression decreased compaction of SKOV-3 spheroids relative to mock.Subsequently, levels of mesenchymal markers such as N-cadherin, snail and fibronectin weresignificantly decreased in Wnt5A/SKOV-3 cells compared to mock. Integrin array analysis showedincreased expression levels of α5, αV and β1 integrins and decreased expression levels of α2, α4,αvβ3, β3 and β4 integrins in transfected cells compared to mock. It is noteworthy that the observedchanges in integrins expression was similar in both monolayer and spheroid models inWnt5A/SKOV-3 cells. Whereas α5β1 serves as fibronectin receptor, correspondingly, here, wefound increased compaction and adhesion of cells in the presence of fibronectin in transfected cellscompared to mock. Wnt5A/SKOV-3 cell migration were increased compared to mock. Interestingly,in the presence of BOX5, we found decreased mRNA levels of ITG A5 and AV integrins as well asreduced levels of Wnt5A/SKOV-3 cell migration compared to mock. Altogether our data suggest thatWNT5A may play a promoting role in EOC progression through modulation of integrin expression.