KLF6在砷诱导肺上皮细胞恶性转化中的作用与机制研究

项目名称： KLF6在砷诱导肺上皮细胞恶性转化中的作用与机制研究

项目编号： No.81502765

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 罗芬

作者单位： 中国人民解放军第三军医大学

项目金额： 18万元

中文摘要： 我国砷污染形势严峻，深入研究砷肺癌发生机制十分迫切。我们发现砷致肺上皮细胞恶性转化中抑癌基因klf6显著降低，基因网络分析显示其处于差异基因核心。p21和cdh1均为细胞恶性转化中关键分子。前期结果及生物信息学提示砷致klf6降低可能通过抑制p21和cdh1，引起肺上皮细胞恶性转化。本研究以砷致肺上皮细胞恶性转化为模型，通过qRT-PCR、免疫印迹技术研究该过程中klf6、p21和cdh1表达情况及关系；利用EMSA、荧光素酶报告基因、ChIP等研究KLF6对p21和cdh1的转录调控；最后利用过表达和RNAi研究klf6对该过程的影响，及p21和cdh1在其中的作用和地位。结果将有助于明确klf6在砷致肺上皮细胞恶性转化中的作用，亦有望阐明抑癌基因klf6与EMT关键开关cdh1之间的调控关系，为防治低剂量砷这一重要环境污染物所致肺癌提供早期 预警靶点和防治线索。

中文关键词： 砷暴露；转录调控；E-钙粘蛋白；Kruppel样因子6；抑癌基因

英文摘要： Arsenic contamination is a very important situation in China, it is urgent to study its effect on lung carcinogenesis. We found cancer suppressor gene klf6 decreased during the arsenic induced lung epithelial cells malignant transformation, bioinformatics analysis indicated that it palys an important role in the gene network. Protein p21 and cdh1 are key players in cells malignant transformation. Our preliminary results suggested arsenic induced klf6 downregulation may induced lung epithelial cells malignant transformation by decreaseing p21 and cdh1 expression. We will using qRT-PCR、Western blotting to study the expression relationship of klf6, p21 and cdh1during arsenic induced lung epithelial cells malignant transformation; and then using EMSA, luciferase reporter assay and ChIP assay to study the regulation of klf6 on p21 and cdh1; After that, we will using over-expression and RNAi technologies to study the role of Klf6 and the dependence of p21 and cdh1in arsenic induced cells malignant transformation. Our results will be helpful to clarify the role and mechanism of klf6 in arsenic induced cells malignant transformation, prove a new direct regulation pathway between klf6 and cdh1, and provide new clues for finding biomarkers for early detection and early prevention of arsenic induced lung cancer.

英文关键词： arsenic exposure;transcriptional regulation;E-cadherin;Kruppel-like factor 6;tumor supressor gene