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Cell Journal، جلد ۱۷، شماره Suppl ۱، صفحات ۲۳-۲۳
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عنوان انگلیسی Ps-6: Wnt5A Promotes Spheroid Formation and Increase Integrinb1 Expression in Human Ovarian Cancer Cell Line; SKOV-3
چکیده انگلیسی مقاله Objective: Metastasis of ovarian cancer (OC) requires formation of OC cell spheroids in peritoneum and their further adherence to mesothelium which is rich in collagens, laminin and fibronectin and requires integrinb1 up-regulation in OC cells. Our previous study showed that Wnt5A influence adhesion and survival of monolayer culture of human OC cell line, SKOV-3. This study sought to determine Wnt5A role in spheroid formation of SKOV-3 cells and its impact on integrinb1 expression. Materials and Methods: Wnt5A was overexpressed or knocked-down in SKOV-3 cells. Cells were stably transfected with pcDNA3.2/V5-DEST-Wnt5A construct or with pcDNA3.2/V5-DEST without Wnt5A (mock) by using Lipofectamine 3000. Wnt5A was knocked down using specific small interfering RNA (siRNA) against Wnt5A, and as negative controls non-target siRNAs (scramble) and transfection reagent (TR, Lipofectamine 2000) were only used. Following transfections, expression of Wnt5A was assessed by real-time qPCR and western blot analysis. Integrinb1 expression was analyzed using real-time qPCR. Spheroids formation ability was evaluated by using hanging drop technique. Migration of Wnt5A overexpressed cells was assessed using scratch assay. Reproductive viability of overexpressed Wnt5A cells was determined by colony formation assay. Results: Wnt5A was increased by 2.4-fold accompanied with increased compaction of spheroids. siRNA Wnt5A transfected cells showed 70% decreased Wnt5A expression compared with scramble or TR which led to decreased spheroid growth and compaction. Migration and colony formation in Wnt5A overexpressed cells were increased compared to mock. In addition, Wnt5A overexpressed cells showed 8.7-fold increased integrinb1 expression compared to mock. Conclusion: Our results suggest that Wnt5A may promote ovarian cancer cells spheroid formation and compaction which requires up-regulation of integrins.
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نویسندگان مقاله v عظیمیان | v azimian


g حسین | g hossein


m ابراهیمی | m ebrahimi


نشانی اینترنتی http://celljournal.org/journal/article/abstract/62
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