【摘要】 目的探讨褪黑素对子宫内膜上皮细胞运动性和上皮间质转化(EMT)的影响,及其对Numb/Notch1/Snail/E-cadherin信号通路的调控作用。方法采用免疫组化法检测20份正常子宫内膜和20份异位子宫内膜中Notch1、Numb、E-cadherin和Snail的表达。使用20nmol/L 17β-雌二醇和5 mmol/L褪黑素处理子宫内膜上皮细胞。使用细胞计数试剂盒-8(CCK-8)评估细胞增殖。使用Transwell实验评估细胞迁移和侵袭能力。Western blot检测相关蛋白表达。结果与正常子宫内膜相比,子宫内膜异位症子宫内膜中的Notch1和Snail的阳性染色评分显著升高,而Numb和E-cadherin显著降低(P<0.05)。培养72 h后,与对照组相比,17β-雌二醇处理组的子宫内膜上皮细胞的增殖能力显著升高(P<0.05)。与17β-雌二醇组相比,褪黑素处理组的细胞增殖能力显著降低(P<0.05)。与对照组相比,17β-雌二醇组的迁移和侵袭细胞数显著升高(P<0.05)。与17β-雌二醇组相比,褪黑素组的迁移和侵袭细胞数显著降低(P<0.05)。与对照组相比,17β-雌二醇组的Numb和E-cadherin的蛋白表达水平显著降低,而Notch1和Snail显著升高(P<0.05)。与17β-雌二醇组相比,褪黑素组的Numb和E-cadherin蛋白表达水平显著升高,而Notch1和Snail蛋白表达水平显著降低(P<0.05)。结论子宫内膜异位症患者的子宫内膜中存在异常的EMT。褪黑素可通过调节Numb/Notch1/Snail/E-cadherin信号通路有效抑制17β-雌二醇诱导的子宫内膜上皮细胞的运动性和EMT。更多还原

【Abstract】 Objective To investigate the effects of melatonin on endometrial epithelial cell motility and epithelial-mesenchymal transition(EMT), and its regulation on Numb/Notch1/Snail/E-cadherin signaling pathway.Methods The expressions of Notch1, Numb, E-cadherin and Snail in 20 samples of normal endometrium and 20 samples of ectopic endometrium were detected by immunohistochemistry. Endometrial epithelial cells were treated with 20 nM 17β-estradiol and 5 mM melatonin. Cell proliferation was assessed using the Cell Counting Kit-8(CCK-8). Transwell assay was used to assess cell migration and invasiveness. Western blot was used to detect the expressions of related proteins.Results Compared with normal endometrium, Notch1 and Snail positive staining scores were significantly increased in endometriosis, while Numb and E-cadherin were significantly decreased(P<0.05). After 72 h of culture, compared with the control group, the proliferation ability of endometrial epithelial cells in the 17β-estradiol-treated group was significantly increased(P<0.05).Compared with the 17β-estradiol group, the cell proliferation ability of the melatonin-treated group was significantly reduced(P<0.05). Compared with the control group, the number of migrating and invading cells in the 17β-estradiol group was significantly increased(P<0.05). Compared with the 17β-estradiol group, the number of migrating and invading cells in the melatonin group was significantly reduced(P<0.05). Compared with the control group, the protein expression levels of Numb and E-cadherin in the 17β-estradiol group were significantly reduced, while Notch1 and Snail were significantly increased(P<0.05). Compared with the 17β-estradiol group, the expression levels of Numb and E-cadherin proteins in the melatonin group were significantly increased, while Notch1 and Snail were significantly decreased(P<0.05).Conclusion There is an abnormal EMT in the endometrium of patients with endometriosis. Melatonin can effectively inhibit 17β-estradiol-induced endometrial epithelial cell motility and EMT by regulating Numb/Notch1/Snail/E-cadherin signaling pathway.更多还原