核转录因子KLF4抑制LILRB2介导JNK磷酸化调控EMT依赖肺腺癌转移的机制研究

肺腺癌是非小细胞肺癌主要的组织学类型，其复发、转移严重影响患者预后。申请人前期证实转录因子KLF4在JNK介导的EMT依赖的肺腺癌转移中起到关键作用。后续TCGA数据生信分析提示LILRB2是转录因子KLF4的靶基因之一，本项目拟在此工作基础上，阐释KLF4调控LILRB2机制，利用ChIP、测序技术确定KLF4 与LILRB2结合位点，利用Pull-down及蛋白质谱技术明确是否有伴侣蛋白存在；同时干扰或者过表达LILRB2观察对肺腺癌细胞侵袭转移能力的影响，以及下游JNK磷酸化及EMT标志物表达的影响，进而激动或抑制JNK活化进行回复验证；进一步利用尾静脉注射肺转移和心脏注射骨转移动物模型获得在体证据；最后在组织样本中检测KLF4/LILRB2/JNK/EMT信号轴各成员的表达，与临床特征做单因素/多因素相关性分析和预后分析，揭示新分子靶点的临床转化意义。

Lung adenocarcinoma is the main histological type of non-small cell lung cancer, and its recurrence and metastasis affect the prognosis of patients. The applicant previously confirmed that KLF4 plays a key role in JNK mediated EMT dependent lung adenocarcinoma metastasis. Subsequent TCGA data analysis showed that LILRB2 was one of the target genes of KLF4. Based on this work, this project intends to explain the mechanism of KLF4 regulating LILRB2, the binding sites of KLF4 and LILRB2 were determined by ChIP and gene sequencing, and the presence of chaperone was determined by pull-down assay and protein mass spectrometry. At the same time, the effect of interference or overexpression of LILRB2 on the invasion and metastasis of lung adenocarcinoma cells, as well as the downstream JNK phosphorylation and EMT marker expression were observed, so as to activate or inhibit JNK for rescue verification. The animal models of lung metastasis by tail vein injection and bone metastasis by heart injection were used to obtain in vivo evidence. Finally, the expression of KLF4 / LILRB2 / JNK / EMT signal axis was detected in human lung adenocarcinoma tissue samples, and univariate / multivariate correlation analysis and survival analysis were performed to reveal the clinical significance of new molecular targets.