Wnt/β-catenin通路抑制剂Wnt-C59抑制子宫内膜异位症子宫内膜干细胞的增殖

Inhibitory effect of Wnt/β-catenin pathway inhibitor Wnt-C59 on endometrial stem cell proliferation in endometriosis

目的分析Wnt/β-catenin通路抑制剂Wnt-C59对子宫内膜异位症(EM)子宫内膜干细胞增殖的影响。方法分离纯化获得子宫内膜干细胞,随机分为对照组、低剂量组(5 mmol/L Wnt-C59)和高剂量组(30 mmol/L Wnt-C59)。MTT法分析各组细胞增殖情况,Western blotting法分析细胞凋亡蛋白及Wnt/β-catenin通路蛋白的表达变化,酶联免疫吸附测定法分析LC3、Beclin1水平变化。结果与对照组比较,低剂量组和高剂量组细胞增殖受到明显抑制,Wnt3a及β-catenin蛋白水平明显降低,细胞凋亡蛋白Caspase-3及Caspase-9水平明显升高,LC3、Beclin1蛋白水平明显升高(P<0.05),且高剂量组均更为明显(P<0.05)。Pearson相关性分析显示,Wnt3a及β-catenin蛋白水平均与细胞增殖抑制率呈负相关(P<0.05)。结论Wnt/β-catenin通路抑制剂Wnt-C59能有效抑制EM子宫内膜干细胞增殖,促进其凋亡、自噬作用,其作用可能与其对Wnt/β-catenin信号通路的抑制有关。

Aim To analyze the impacts of Wnt/β-catenin pathway inhibitor Wnt-C59 on the proliferation of endometrial stem cell in endometriosis(EM).Methods Endometrial stem cell were separated and purified,and then randomly divided into control group,low dose group(5 mmol/L Wnt-C59)and high dose group(30 mmol/L Wnt-C59).MTT assay was used to analyze the proliferation of cell in each group.Western blotting assay was used to analyze the expressions of apoptotic protein and Wnt/β-catenin pathway protein.Enzyme-linked immunosorbent assay was used to analyze the changes of LC3 and Beclin1 levels.Results Compared with the control group,the proliferation of endometrial stem cell was significantly inhibited,the levels of Wnt3 a andβ-catenin proteins were significantly down-regulated,while apoptotic proteins Caspase-3 and Caspase-9 were significantly up-regulated and LC3,Beclin1 proteins levels were significantly up-regulated in low dose group and high dose group(P<0.05),besides differences above it was more significant in the high dose group(P<0.05).Pearson correlation analysis showed that Wnt3 a andβ-catenin levels were negatively correlated with the inhibition rate of cell proliferation(P<0.05).Conclusion Wnt/β-catenin pathway inhibitor WntC59 can effectively inhibit the proliferation of endometrial stem cell in EM,boosting the apoptosis and autophagy of endometrial stem cell,and its impact may be related to its effect of inhibiting Wnt/β-catenin pathway.