ATF3通过与JunB相互作用负向调控炎症和子宫内膜癌进程的机制研究

Existing evidence indicates that PPI (protein-protein interaction) contributes to the tumorigenesis of endometrial carcinoma, but the mechanisms involved in it remain largely unknown. In our previous study, we found that ATF3 participating in the development of various different kinds of cancer, expressed higher in the epithelium of normal endometrial tissue than that detected in pathnological tissue, and interacted with JunB which expressing higher in the epithelium of endometrial cancer tissue than that detected in normal endometrial tissue, via yeast-two-hybrid assay. Additionally, over-expression of ATF3 led to the inhibition of cell proliferation, migration and invasion, with the down-regulation of JunB and inflammatory factor. Therefore, we put forward a hypothesis that ATF3 might inhibit the expression of inflammatory genes and the development of endometrial cancer by binding with JunB andmitigating with it. This study was designed to utilize the mRNA expression microarray analysis, CRISPR/Cas9 and orthotopic endometrial carcinoma model in nude mice combined with molecular biological techniques to explore the cell signal transductions of ATF3 and JunB in EC, demonstrate the mechanism of development of EC, and provide new therapeutic targets for it, which contributes greatly to the patients suffering from endometrial cancer.

蛋白质相互作用在子宫内膜癌的发病及进程中起重要作用，但其具体机制尚不清楚。前期工作发现，广泛参与肿瘤发生与进程的转录因子ATF3在内膜癌组织中上皮细胞内的表达低于正常内膜组织中上皮细胞内的表达；通过酵母双杂交技术在正常人子宫内膜组织中筛选出的、可与ATF3结合并相互作用的转录因子JunB在内膜癌组织上皮细胞内高表达，正常内膜组织上皮细胞内低表达；过表达ATF3后癌细胞增殖、侵袭、迁移能力减弱，同时JunB、炎症因子表达下调。由此我们提出一个假设，ATF3通过与JunB相互作用下调JunB表达，抑制炎症基因表达及内膜癌进程。本项目拟通过表达谱芯片技术筛查两者结合启动的下游信号通路，通过分子生物学技术验证，诠释发挥作用的具体机制，并应用CRISPR/Cas9和原位移植裸鼠模型结合的方法，构建JunB基因敲除细胞和动物模型，为治疗内膜癌提供新治疗策略，为临床提供新治疗靶点，具有显著社会效益和意义

子宫内膜癌是女性生殖系统最常见的恶性肿瘤。蛋白质相互作用在子宫内膜癌的发病及进程中起重要作用，但其具体机制尚不清楚。课题组预实验发现，广泛参与肿瘤发生与进程的转录因子ATF3在内膜癌组织中上皮细胞内的表达低于正常内膜组织中上皮细胞内的表达。课题组在预实验的基础上，通过酵母双杂交技术在正常人子宫内膜组织中筛选出可与ATF3结合并相互作用的转录因子JunB，并发现ATF3通过与JunB相互作用下调JunB表达，抑制炎症基因表达及内膜癌进程。本项目通过表达谱芯片技术筛查两者结合启动的下游信号通路，通过分子生物学技术验证，诠释了发挥作用的具体机制，并应用CRISPR/Cas9技术，构建了JunB基因敲除的子宫内膜癌细胞模型，在理论上加深了我们对内膜癌复杂的发病机制的理解,为治疗内膜癌提供了新的治疗策略。与此同时，作为课题的延伸之一，我们发现与ATF3存在相互作用的转录因子SOX3促进上皮性卵巢癌细胞的增殖和转移；作为课题的延伸之二，我们利用CRISPR技术对转录因子进行了高通量筛选并对其进行了功能学验证。我们的研究进一步阐明了转录因子及其相互作用在细胞模型、疾病尤其是女性生殖系统恶性肿瘤发病中的重要作用，为寻找新治疗靶点提供了理论和实验依据。