| Abstract:
Introduction: Ovarian cancer is the fifth leading cause of death from cancer in women and is the leading cause of death from gynecological cancer. Several reports showed the involvement of Wnt signaling alteration in different type of cancers and recently increased expression of Wnt5a was reported in epithelial ovarian cancer. This study sought to determine the effect of Wnt5a over-expression on cell cycle and survival.
Materials and Methods: Human Wnt5a (hWnt5a) was cloned, and inserted into mammalian vector pIRES-EGFP which harbors CMV promoter.
OVCAR3 and SKOV3 epithelial ovarian carcinoma cell lines were transfected with vector harboring hWnt5a by using Fugene HD lipofection kit, non-transfected and mock cells were used as controls. In order to select the stabled transfected cells, G418 antibiotic was used in a proper dose for one month in the culture media. After getting stabled transfected, cells RT-PCR and Immunocytochemistry (ICC) were performed for detection of hWnt5a expression in the ovarian cancer cell lines. Furthermore, MTT assay, and scratch healing test were used to assess modulation of cell proliferation and cell migration for 12, 24 and 48 hours of cell culture in the absence of serum.
The assessment of cell cycle was done by using flowcytometric assay after 12 hr, 24 hr and 48 hr culture.
Results: RT-PCR and ICC assays showed strong expression of hWnt5a in transfected ovarian cancer cell lines. Transfected cells showed significant increase in cell survival compared to controls in both cell lines, as revealed by MTT assay. Scratch healing assay showed increased motility of both transfected cell lines compared to controls. However, cell cycle analysis did not revealed significant differences between transfected cells and controls.
Conclusion: Our data shows that hWnt5a increase cell survival and motility of ovarian cancer cell lines, which would suggest implication of Wnt5a in metastasis and ovarian cancer progression.
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