缺氧炎症微环境中肝癌细胞上皮间质转化相关基因甲基化的变化及作用机制

项目名称： 缺氧炎症微环境中肝癌细胞上皮间质转化相关基因甲基化的变化及作用机制

项目编号： No.81201584

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 肿瘤学1

项目作者： 井莹莹

作者单位： 中国人民解放军第二军医大学

项目金额： 23万元

中文摘要： 肝癌高转移复发的特性是影响其远期疗效的主要瓶颈。肿瘤的转移复发与微环境密切相关，局部缺氧及炎症是肿瘤微环境最显著的两个特点。我们前期研究发现：缺氧和炎症因子都能明显促进肝癌细胞上皮间质转化（EMT），而缺氧和炎症又可以调控肿瘤细胞的DNA甲基化，但缺氧联合炎症因子对肝癌细胞EMT相关基因甲基化的影响目前仍未明确。本课题拟在这些线索的基础上，采用Transwell、划痕实验及裸鼠肝癌转移模型等方法，研究缺氧联合炎症因子对肝癌细胞EMT及侵袭转移能力的影响；用高通量DNA甲基化芯片筛选肝癌细胞EMT相关的差异基因，并采用MSP鉴定，探讨缺氧联合炎症因子对肝癌细胞EMT相关基因甲基化调控的分子机制；结合临床标本，进一步研究肝癌组织中筛选出的甲基化位点与临床预后的相关性。本研究结果不但可以加深对肿瘤微环境影响肿瘤转移复发机制的理论认识，同时将为探索针对微环境的抗肿瘤转移复发新策略研究提供实验基础。

中文关键词： 肝癌；上皮间质转化；微环境；转移；侵袭

英文摘要： High metastatic recurrence characteristics of hepatoma affect the long-term treatment. Tumor recurrence and metastasis is closely related to the tumor microenvironment, which has two significant characteristics: hypoxia and inflammation. Our previous study showed that: hypoxia and inflammatory factors could significantly promote the epithelial-mesenchymal transition (EMT) of hepatoma cells. Hypoxia and inflammation can regulate DNA methylation of tumor; however, the effect of hypoxia combined with inflammation factor on EMT related-gene methylation in hepatoma cell is not yet clear. On the basis of these clues, this topic is intended to study the effect of hypoxia combined with inflammation factor on hepatoma EMT as well as invasion and metastasis by using transwell, wound healing assay and nude mouse tumor metastasis model; EMT related-gene methylation is detected by high-throughput DNA methylation microarray and confirmed by MSP, which to explore the molecular mechanism of EMT related-gene methylation regulated by hypoxia combined with inflammation factor; Further study the relation of screened methylation sites and clinical prognosis in the clinical specimens. The results of this study cannot only deepen our understanding of tumor microenvironment and tumor metastasis, recurrence, but also provide the exper

英文关键词： hepatoma；epithelial mesenchymal transformation (EMT) ；microenvironment；metastasis；invasion