子宫腺肌病子宫内膜间质细胞中14-3-3σ表达及启动子甲基化异常的研究

Study on 14-3-3σ expression in interstitial cells of endometrium in patients with adenomyosis and abnormal promoter methylation

目的:通过研究子宫腺肌病间质细胞中14-3-3σ基因表达及启动子区CpG岛甲基化水平,探讨14-3-3σ基因在子宫腺肌病发生和发展中的作用及意义。方法:分离、纯化培养人原代子宫腺肌病子宫内膜间质细胞,同期分离纯化卵巢良性肿瘤患者原代子宫内膜间质细胞为对照细胞。采用Western-Blotting和RT-PCR方法分别检测并比较14-3-3σ的蛋白及mRNA表达水平;采用MSP方法检测并比较14-3-3σ基因启动子区CpG岛的甲基化水平。结果:在子宫腺肌病间质细胞中,14-3-3σ的蛋白及mRNA表达水平均显著低于对照细胞。MSP结果显示,其基因启动子区CpG岛存在甲基化。结论:子宫腺肌病间质细胞中,14-3-3σ蛋白及mRNA表达下调,可能是由于其基因启动子区CpG岛异常甲基化所引起的。14-3-3σ基因启动子甲基化及表达异常可能参与了子宫腺肌病的发病过程。

Objective： To explore the role and significance of 14 -3 -3σ gene in occurrence and aggravation of adenomyosis by researching 14 - 3 - 3σ gene expression and promoter methylation of CpG island in interstitial cells of endometrium in patients with adenomy- osis. Methods： Human primary interstitial ceils of endometrium in patients with adenomyosis were isolated and purified, purified primary interstitial cells of endometrium in patients with benign ovarian tumors were isolated during the same period as control cells. Western - Blot- ting and RT - PCR were used to detect and compare the expression levels of 14 -3 -3σ protein and mRNA; MSP was used to detect and compare methylation level of CpG island in 14 -3 -3σ gene promoter. Results： The expression levels of 14 -3 -3σ protein and mRNA in interstitial cells of endometrium in patients with adenomyosis were significantly lower than those in control cells ; MSP results showed that CpG island in 14 -3 -3or gene promoter was methylated. Conclusion： 14 -3 -3σ protein and mRNA are down -regulated in interstitial cells of endometrium in patients with adenomyosis, which may be induced by abnormal methylation of CpG island in 14 -3 -3σ gene promoter; 14 -3 -3σ gene promoter methylation and abnormal expression may be involved in pathogenesis of adenomyosis.