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目的检测环氧化酶-2(COX-2)基因启动子区CRE位点甲基化状况,探讨其在子宫内膜异位症(EMs)发生中的作用。方法收集南昌大学第一附属医院40例子宫内膜异位症患者的子宫内膜作为病例组,同期40例正常子宫内膜作为对照组,采用焦磷酸盐测序技术检测病例组和对照组子宫内膜组织中COX-2基因启动子区CRE位点甲基化频率;采用实时定量PCR及免疫组化技术分别检测病例组和对照组子宫内膜组织中COX-2 mRNA及蛋白的表达;分析COX-2基因启动子区CRE位点甲基化频率与COX-2表达水平的相关性。结果对照组内膜组织COX-2基因启动子区CRE位点的甲基化频率(40.81%±6.10%)显著高于病例组(14.74%±2.84%),P<0.05;而对照组COX-2 mRNA表达水平(0.98±0.37)及蛋白表达水平(1.00±0.73)显著低于病例组的mRNA表达水平(2.63±0.44)及蛋白表达水平(2.74±1.18),P均<0.05。随着COX-2基因启动子区CRE位点甲基化频率升高,COX-2基因表达水平下降,二者呈负相关(P<0.05)。结论 COX-2基因启动子区CRE位点去甲基化与EM s患者在位内膜COX-2高表达相关,在EM s的发生、发展中具有重要作用,提示子宫内膜异位症是一种表观遗传性疾病。
Objective To investigate the methylation status of CRE site in COX-2 gene promoter region and its role in the pathogenesis of endometriosis (EMs). Methods 40 cases of endometriosis in the First Affiliated Hospital of Nanchang University were collected as the case group and 40 cases of the normal endometrium as the control group. Pyrosequencing was used to detect the intrauterine in the case group and the control group COX-2 gene promoter region of the membrane methylation frequency of CRE sites; using real-time quantitative PCR and immunohistochemical techniques were detected in the case group and the control group, the endometrial COX-2 mRNA and protein expression; analysis of COX -2 Gene Promoter Region CRE Site Methylation Frequency and COX-2 Expression Level Correlation. Results The methylation frequency of CRE site in COX-2 gene promoter region (40.81% ± 6.10%) in the control group was significantly higher than that in the case group (14.74% ± 2.84%, P <0.05) 2 mRNA expression level (0.98 ± 0.37) and protein expression level (1.00 ± 0.73) were significantly lower than those in case group (2.63 ± 0.44) and protein expression level (2.74 ± 1.18), P <0.05. With the increase of methylation frequency of CRE site in COX-2 gene promoter region, the expression of COX-2 gene decreased, and the two were negatively correlated (P <0.05). Conclusions The demethylation of COX-2 promoter in CRE promoter region is associated with the high COX-2 expression in eutopic endometrium of EM s patients, which plays an important role in the development and progression of EM s, suggesting that endometriosis is An epigenetic disorder.