TMEM196基因在化学致肺癌过程中的DNA甲基化调控和功能研究

项目名称： TMEM196基因在化学致肺癌过程中的DNA甲基化调控和功能研究

项目编号： No.81202238

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 预防医学、地方病学、职业病学、放射医学

项目作者： 刘文斌

作者单位： 中国人民解放军第三军医大学

项目金额： 24万元

中文摘要： TMEM196基因是申请者前期在研究化学物致肺癌过程中筛选出的高甲基化基因。进一步研究发现，该基因在大鼠和人肺癌组织以及多种肺癌细胞株中均发生高甲基化且表达缺失，而在正常肺组织中未发生甲基化且高表达，提示该基因受DNA甲基化调控，可能在癌变过程中发挥重要作用。经文献检索，尚未见有关该基因表达调控机制和功能的报道。本研究拟在前期建立化学致肺癌模型的基础上，检测癌变各阶段TMEM196基因甲基化、mRNA和蛋白表达水平，结合去甲基化药物处理实验，阐明该基因在化学致肺癌过程中的DNA甲基化调控机制；构建该基因过表达和沉默细胞模型，结合体内裸鼠成瘤实验，从细胞增殖、凋亡、周期、迁移等方面分析该基因在肺癌发生中的主要生物学功能，并在化学致癌模型中进一步验证。本项目不仅对阐明化学致癌和肺癌发病机制具有重要意义，而且为深入研究TMEM196基因在信号通路、生长发育、疾病发生等方面的作用奠定了坚实基础。

中文关键词： TMEM196基因；化学致癌；DNA 甲基化；肺癌；表观遗传调控

英文摘要： TMEM196 gene is a hypermethylated gene screened previously by applicant in the process of research about chemical-induced lung cancer. Further research found that promoter methylation and loss of TMEM196 gene expression occur in chemical-induced rat lung cancer tissues, human lung cancer tissue and a series of cell lines. However, this gene with unmethylated express in normal lung tissues. Results of previous research suggest that this gene regulated by DNA methylation may play an important role during lung carcinogenesis. However, related research about TMEM196 gene expression regulation and function has not yet been reported in the literature. On the basis of chemical-induced rat lung cancer model established previously, this study will detect the status of TMEM196 gene methylation, mRNA and protein expression during rat lung carcinogenesis, combined with demethylation drug treatment experiment, to clarify DNA methylation regulation mechanism for TMEM196 gene expression during chemical-induced lung carcinogenesis. Then, through construction of TMEM196 gene expression reconstruction and silence cell model combined with nude mouse tumorigenicity assays in vivo, we will explore the biological functions of the gene in cells growth, apoptosis, cycle, migration, and further validate in chemical-induced animal model.

英文关键词： TMEM196 gene；Chemical carcinogenesis；DNA methylation；Lung cancer；Epigenetic regulation