Objective. To investigate the effects of Cyclocarya paliurus polysaccharide (CPP) on the proliferation, migration, invasion, and apoptosis of human glioma U251 cells and further explore the underlying mechanism. Methods. U251 cells were cultured in vitro and treated with various concentrations (25, 50, 75, 100, 125, and 150 μmol/L) of CPP for 24, 48, and 72 h. Cell counting kit-8 was used to detect the activity of cell proliferation. Wound-healing assay, Transwell assay, and flow cytometry were used to measure the effects of CPP on the migration, invasion, and apoptosis of U251 cells, respectively. Western blotting was used to determine the protein expression involved in the GSK3β/β-catenin signaling pathway and its downstream genes related to proliferation, migration, invasion, and apoptosis including Cyr61, CCND1, Vimentin, and Slug. Meanwhile, qRT-PCR was used to detect the mRNA levels of Cyr61, CCND1, Vimentin, and Slug. Results. We found that CPP not only could inhibit the proliferation, migration, and invasion of U251 cells but also promote its apoptosis in vitro. Besides, CPP could significantly inhibit the phosphorylation and decrease the protein levels of GSK3 β at ser9 site (), and thus increasing the phosphorylation of β-Catenin at ser33/37 site (), resulting in β-Catenin degradation. In addition, we also found that CPP could downregulate the mRNA () and protein expression () of downstream genes of GSK3 β/β-catenin signaling pathway including Cyr61, CCND1, Vimentin, and Slug, which are related to proliferation, migration, invasion, and apoptosis. Conclusion. CPP could inhibit the expression of GSK3β, promote the degradation of β-catenin, and downregulate the levels of GSK3β/β-catenin downstream genes including Cyr61, CCND1, Vimentin, and Slug, which regulate the proliferation, migration, invasion, and apoptosis of glioma cells.

中文翻译：

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Cyclocarya paliurus多糖通过GSK3β/β-Catenin信号通路抑制胶质瘤细胞U251的增殖，迁移和侵袭并促进细胞凋亡。

目标。为了研究环青冈多糖（CPP）对人胶质瘤U251细胞增殖，迁移，侵袭和凋亡的影响，并进一步探讨其潜在机制。方法。U251细胞在体外进行培养，并用不同浓度的（25，50，75，100，125和150处理过的 μ摩尔/ L）的CPP 24,48，和72小时。细胞计数试剂盒-8用于检测细胞增殖的活性。伤口愈合测定，Transwell测定和流式细胞仪分别用于测量CPP对U251细胞迁移，侵袭和凋亡的影响。Western印迹被用于确定参与GSK3蛋白表达β / β-catenin信号通路及其下游基因与增殖，迁移，侵袭和凋亡相关，包括Cyr61，CCND1，Vimentin和Slug。同时，使用qRT-PCR检测Cyr61，CCND1，Vimentin和Slug的mRNA水平。结果。我们发现CPP不仅可以抑制U251细胞的增殖，迁移和侵袭，而且可以促进其体外凋亡。此外，CPP能显著抑制磷酸化和减少GSK3蛋白水平β在ser9网站（），从而增加在ser33 / 37位点的β-连环蛋白的磷酸化（），导致β-连环素降解。此外，我们还发现CPP可以下调mRNA（）和蛋白质表达（） GSK3的下游基因的β / β连环蛋白信号传导途径，包括的Cyr61，CCND1，波形蛋白，和蛞蝓，其涉及到细胞增殖，迁移，侵入和细胞凋亡。结论。CPP可以抑制GSK3的表达β，促进的降解β连环蛋白，并下调GSK3水平β / β连环蛋白的下游基因，包括的Cyr61，CCND1，波形蛋白，和蛞蝓，其调节细胞增殖，迁移，侵袭和胶质瘤细胞凋亡。