17β-雌二醇对子宫内膜异位症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响

目的研究17β-雌二醇(17β-E2)对子宫内膜异位症(内异症)患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响,探讨Wnt/β-catenin信号通路在介导雌激素促进内异症发生发展的作用。方法体外分离培养内异症患者在位子宫内膜间质细胞。用不同浓度17β-E2处理子宫内膜间质细胞48 h;此后选用10-10mol/L 17β-E2处理子宫内膜间质细胞12、24和48 h,逆转录聚合酶链反应(RT-PCR)和免疫印迹法(Western blotting)检测17β-E2处理前后子宫内膜间质细胞β-catenin mRNA和蛋白的表达水平。同法分析雌激素受体拮抗剂ICI182,780(10-6mol/L)对17β-E2促进β-catenin mRNA和蛋白表达的影响。免疫组织化学染色观察17β-E2作用后β-catenin在子宫内膜间质细胞中的定位。结果17β-E2能明显促进内异症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白的表达,并呈剂量和时间依赖性,于10-10mol/L作用48 h最明显。雌激素受体拮抗剂ICI182,780能明显抑制17β-E2对子宫内膜间质细胞β-catenin mRNA和蛋白的表达。免疫组织化学染色发现17β-E2能促进β-catenin在子宫内膜间质细胞核内的表达。结论雌激素可能通过激活Wnt/β-catenin信号通路促进内异症在位子宫内膜的异位种植。

Treatment of 17β-estradiol on expression of β-catenin mRNA and protein in cultured entopic endometrial stromal cell of patients with endometriosis

Objectives: To investigate the expression of β-catenin mRNA and protein,the protein localization within the cultured entopic endometrial stromal cell(ESC) of endometriosis by estrogen treatment,and the role of Wnt signaling pathways in pathogenesis of endometriosis.Methods: Entopic ESCs of endometriosis patient were separated and cultured.The cultured cells were treated by 17β-estradiol(17β-E2) at various concentrations for 48h or 10-10 mol/L 17β-E2 for different durations(0,12,24,48h).The expressions of β-catenin mRNA and protein in ESC of endometriosis were detected by RT-PCR and western-blotting.Meanwhile,the expression of β-catenin mRNA in the cells treated by 17β-E2 plus ER inhibitor ICI 182,780(10-6 mol/L) was detected by the same method.Localization of β-catenin protein in the cells was observed after 17β-E2 treatment for 48h by immunocytochemistry.Results: RT-PCR and western-blotting showed that 17β-E2 stimulated the expression of β-catenin mRNA and protein in ESC of endometriosis in a time-and dose-dependent manner,and the effect was maximal at 10-10 mol/L for 48 h treatment.The 17β-E2 inhibitor ICI182,780 significantly reduced the expression of β-catenin mRNA and protein in ESC treated by 17β-E2.17β-E2 promoted the expression of β-catenin protein in nucleus. Conclusions: It is suggested that the promoted ectopic implantation of the endometriotic endometrium by estrogen involves the activation of Wnt/β-catenin signaling pathways.