雌激素通过ER-α调控的相关差异表达基因在宫颈癌发病过程中的机制研究

The results from the previous and our studies suggested that estrogen and ER-α might play an important role in the pathogenesis of cervical cancer, and its underlying pathogenic mechanisms remain largely to be elucidated. Therefore, we assume that estrogen plays genotype effect mediated by ER-α, which might cause changes of microenvironment or aberrant signaling pathways, and lead to changes of downstream gene expression or activity, affect the biological features of cells, then promote the occurrence and development of cervical cancer. In our previous work, we found that ER-α was highly expressed in cervical cell lines SiHa and HCC94, and we detected 238 direct binding sites of ER-α via Chip-seq technique, approximately 2% of which located in the promoter regions, here we intend to combine with RNA-seq, searching for differentially expressed genes and then combined with the data of Chip-seq, to find out the potential target genes regulated by ER-α. Then we would select a representative high expression gene X, establish pSiliencer-X cell lines by RNAi technique, evaluate its function by cell functional assays in vitro and in vivo. These results will help us to better understand the role of estrogen and ER-α in the pathogenesis of cervical cancer, and provide valuable guidance and references for the patients with cervicer cancer.

结合前人及自身研究基础，我们假设雌激素通过ER-α介导其基因型效应，可能引起微环境或协同信号网络发生异常，导致某些重要靶基因表达或活性改变，使细胞生物学功能发生改变，进而影响其相关信号通路活性发生改变，促进宫颈癌的发生发展。本课题前期研究发现宫颈癌SiHa和HCC94细胞中ER-α高表达，ChIP-seq检测到与ER-α直接结合的基因位点238个，其中结合在启动子区的基因为2%。拟结合RNA-seq，筛选出受ER-α直接调控的差异表达基因，并将RNA-Seq和ChIP-seq数据相结合，找出这两组数据中共同存在的基因，作为ER-α潜在调控靶基因；选取其中典型差异高表达基因X，采用RNAi干扰技术，建立pSilencer-X细胞株，检测其细胞生物学功能，并在整体动物水平验证其作用。研究结果将有助于我们更好理解雌激素及ER-α在宫颈癌发生发展中的作用。

宫颈癌的发生发展过程中雌激素扮演着重要的角色。ER-α作为雌激素的主要核受体之一，在人类生殖系统中高表达。结合前人及自身研究基础，我们假设雌激素通过其核受体ER-α介导其基因型效应，可能引起微环境或协同信号传导网络发生异常，导致某些重要靶基因表达或活性改变，使细胞的生物学功能发生改变，进而影响其相关信号通路活性发生改变，促进宫颈癌的发生发展。本课题研究发现SiHa/HeLa细胞中ER-α为高表达，ChIP-seq检测到SiHa细胞中与ER-α直接结合的基因位点238个，其中结合在启动子区的基因为2%。E2处理SiHa细胞后RNA-seq技术筛选出差异表达基因，并绘制相关促宫颈癌发生的异常信号通路网络；结合ChIP-seq和RNA-seq数据，及宫颈癌临床特征，发现了启动子区能与ESR1直接结合的差异表达基因C3G。本课题的研究结果表明，雌激素可以通过其核受体调控C3G的mRNA及蛋白表达发生变化，使细胞的生物功能发生改变，进而导致宫颈癌的发生发展。