酚类环境雌激素在子宫肌瘤发生发展中的作用和分子机制研究

The pollution of phenolic environmental estrogens is increasingly serious. Small sample epidemiological surveys have revealed their relationship with uterine fibroids. Yet the exact function and molecular mechanism in the development of uterine fibroids remain unclear. On the basis of previous investigation of environmental risk factors and susceptibility biomarker research, this study, focusing on large sample population as the research object and combining internal and external exposure, aims at looking into and analyzing the correlation between phenolic environmental estrogens and uterine fibroids risks. Meanwhile, a polymorphic study on the enzymes involved in the synthesis and metabolism of phenolic environmental estrogens is carried out in order to define the effect and molecular mechanisms of environmental estrogens. Furthermore, by reaching on the effect of phenolic environmental estrogens in the development of uterine fibroids via established uterine fibroid cells model, with the help of RNA-seq and ChIP-seq technologies, the alterations of gene expression pattern and histone modification in uterine fibroid cells caused by phenolic environmental estrogens are detected; The map of uterine fibroid cell genetic and epigenetic variation induced by phenolic environmental estrogen is created in order to analyze the effect of.critical signaling pathways, which helps to screen sensitive biomarkers and medication targets so as to provide theoretical bases for the environmental intervention as well as the prevention of uterine fibroids occurrence.

酚类环境雌激素的环境污染日趋严重，小样本流行病学调研显示其与子宫肌瘤的发病密切相关，但其在子宫肌瘤发生发展中的确切功能和分子机制目前仍不清楚。本研究在前期环境危险因素调查及易感生物标志研究的基础上，以大样本人群为研究对象，结合内暴露和外暴露深入调查分析酚类环境雌激素与子宫肌瘤发病风险的相关性；同时开展酚类环境雌激素相关合成及代谢酶基因多态性研究，从而明确酚类环境雌激素在子宫肌瘤发生发展中的作用和分子机制；构建原代子宫肌瘤细胞模型，研究酚类环境雌激素在子宫肌瘤发生发展中的功能，并结合RNA-seq和ChIP-seq技术，在全基因组水平检测酚类环境雌激素在子宫肌瘤细胞中引起的基因表达和组蛋白修饰的改变，绘制酚类环境雌激素诱发子宫肌瘤细胞遗传及表观遗传变异图谱，分析关键信号通路的影响，筛选敏感的生物标志和药物靶点，为指导环境干预和子宫肌瘤的防治提供理论依据。

子宫肌瘤是女性生殖系统中最为常见的子宫平滑肌良性肿瘤，在35岁以上的女性中子宫肌瘤的发病率高达20%，而在40-50岁妇女中的发病率甚至高达51%-60%且呈逐年上升趋势，是妇科因病切除子宫最常见的病因。因此，深入研究子宫肌瘤的发病原因和发病机制对于预防和治疗子宫肌瘤具有非常重要的意义。在前期研究的基础上，扩大样本量进一步进行流行病学研究检测和子宫肌瘤患者尿液和血液中酚类环境雌激素暴露水平，进一步探讨了酚类环境雌激素与子宫肌瘤发病风险的相关性。我们前期研究证实酚类雌激素中常见的双酚A(BPA)和壬基酚(NP)体外促进子宫肌瘤细胞增殖。本研究中，体外细胞CCK8实验和流式细胞周期实验结果均显示酚类环境雌激素促进人子宫肌瘤细胞增殖。体内动物实验通过构建NCG小鼠人子宫肌瘤皮下移植瘤模型，BPA灌胃组和NP灌胃组小鼠皮下瘤组织体积增长大于对照组。RNA测序联合ChIP测序分析结果显示，细胞增殖和细胞周期相关基因上调，其中关键基因有E2F1, CCND1, CCNE2，IL6，CXCL8等。KEGG信号通路富集分析显示细胞周期，PI3K-AKT、TGF-β信号通路激活。q-PCR及western blot实验证实了以上结果。本项目阐明了酚类环境雌激素促进子宫平滑肌瘤细胞的增殖和细胞周期进程，加速了细胞周期从G0 / G1期向S期的转变，其中关键基因有E2F1,CCND1,CCNE2，IL6等。此外，作为一种外部有害刺激物，酚类雌激素可促进子宫平滑肌瘤中炎症因子的上调。本项目中结合RNA-seq结合和CHIP-seq数据，深入分析酚类环境雌激素通过表观遗传调控对关键基因表达的影响，分析关键信号通路的影响，筛选敏感的生物标志和药物靶点，为临床用于预防及治疗子宫肌瘤提供实验基础和分子标志记物。