缺氧诱导肺癌上皮细胞-间质转化中缺氧诱导因子调控关键靶基因筛选鉴定及其表观遗传学调控研究

The epithelial-to-mesenchymal transition (EMT) promoted by hypoxia, glycolysis and hypoxia inducible factor (HIF) is a key process for cancer metastasis, in which epigenetic regulation plays an essential role. We previously established a EMT model in A549 lung cancer cell line and demonstrated that histone modifications were involved in its cellular trans-differentiation and metastasis. Due to the tight correlation among hypoxia, HIF regulation and DNA modifications (methylation and hydroxymethylation), we aim to collect A549 cells from different time points of hypoxia-induced EMT, profile their HIF binding targets and DNA methylation status by ChIP-BS-Seq, detect both their DNA methylation and hydroxymethylation profiles by HMST-Seq and analyze the regulation of DNA modifications on target binding activity of HIF, and finally perform function analysis on key regulator genes. This study will pave a road for elucidating gene activation program and partial regulation mechanism of EMT, screening new EMT biomarks and potential diagnostic and therapeutic targets of cancer metastasis

由缺氧微环境、糖酵解代谢与缺氧诱导因子（HIF）累积共同促发的癌上皮细胞向间质细胞转化（EMT）过程是形成癌症转移的关键；而表观遗传修饰在其中起到关键调控作用。本课题组此前成功构建了A549肺癌细胞系EMT模型，发现组蛋白修饰与肺癌细胞的转分化和转移潜能紧密相关。鉴于缺氧和HIF调控与DNA修饰（包括甲基化和羟甲基化）的紧密联系，本项目拟利用自主研发的高通量技术，选取缺氧诱导EMT不同阶段的A549细胞，在全基因组范围以ChIP-BS-Seq检测其转录因子HIF结合区序列及甲基化，以HMST-Seq检测所有CCGG位点的甲基化和羟甲基化。通过筛查EMT进程中不同阶段HIF所启动的靶基因谱，两种DNA修饰变化以及其对HIF与靶基因结合的影响，进而鉴定关键基因功能，本研究将为解析癌细胞EMT过程的基因激活程序及部分调控机制，EMT新分子标记物和潜在的癌症转移诊断和治疗靶点等奠定基础