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Sphingomyelin synthase 2 regulates the malignant biological behaviors of ovarian cancer TOV-21G cells through Wnt/β-catenin pathway
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Abstract:
Objective: To investigate whether sphingomyelin synthase 2 (SMS2) regulates the proliferation, migration, invasion and apoptosis of ovarian cancer (OC) TOV-21G cells through Wnt/β -catenin pathway and its mechanism. Methods: The cancer and para-cancerous tissue samples of 21 OC patients diagnosed in the Third Hospital of Wuhan from July 2022 to May 2023 were collected, and the SMS2 expression in the collected tissues was detected by immunohistochemistry. TOV-21G cells were cultured in vitro and grouped into control group, shRNA lentivirus negative control group (sh-NC group), SMS2 shRNA lentivirus group (sh-SMS2 group), Wnt/β-catenin pathway activator LiCl group (LiCl group), and sh-SMS2+LiCl group. Edu staining, Transwell method and flow cytometry were used to detect the proliferation, migration, invasion, and apoptosis of cells in each group. WB assay was used to detect the expression of SMS2, Ki67, cyclin D1, BAX, c-caspase3, Bcl-2 and Wnt/β -catenin pathway-related proteins (β -catenin, c-Myc, MMP-9) in the cells of each group. A TOV-21G cell transplanted tumor model was established in nude mice to observe the effects of SMS2 knockdown on the growth of transplanted tumors and the expression of SMS2 and β-catenin. Results: Compared with the para-cancerous tissues, the expression of SMS2 in OC tissues was increased obviously (P<0.05). After transfection with sh-SMS2, the expression level of SMS2 in TOV-21G cells was decreased obviously (P<0.05), the proliferation, migration, and invasion abilities of TOV-21G cells as well as the protein expression of Bcl-2, β-catenin, c-Myc, and MMP-9 were decreased obviously (all P<0.05), and the cell apoptosis rate, the protein expression of BAX and c-caspase3 were obviously increased (all P<0.05). LiCl could reverse the inhibitory effects of SMS2 knockdown on the proliferation, migration, invasion, and Wnt/β -catenin pathway of TOV-21G cells (all P<0.05). In vivo tumor formation experiments showed that SMS2 knockdown inhibited tumor growth and the expression of SMS2 and β-catenin (all P<0.05). Conclusion: SMS2 knockdown inhibits the proliferation, migration and invasion and promotes apoptosis of OC TOV-21G cells through Wnt/β-catenin signaling pathway, while LiCl treatment can reverse the inhibitory effects of SMS2 knockdown on the proliferation, migration and invasion of TOV-21G cells.
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