NF-κB诱导YAP磷酸化调控mTOR通路参与胃黏膜炎症恶性转化分子机制研究

胃黏膜肠上皮化生及上皮内瘤变为胃癌前病变，是慢性胃炎转化成胃癌或被治愈的关键节点，但具体作用机制不明。项目组前期研究发现：炎症因子TNF-α可直接或通过NF-κB激活YAP促进胃黏膜炎症恶性转化，干扰YAP可显著抑制此事件，提示YAP在致癌变中具重要地位。IRF3是YAP正调控因子，靶向抑制IRF3/YAP能显著抑制胃癌移植瘤生长。故推测炎症因子通过磷酸化YAP以及mTOR通过自噬调控IRF3/YAP促进胃黏膜炎症恶性转化。本研究拟构建胃黏膜炎症恶性转化细胞系与PDX小鼠模型，用慢病毒干扰、染色质免疫沉淀等方法，在转录水平、蛋白水平及翻译后修饰水平研究Hippo-YAP、mTOR-YAP及NF-κB/IRF3-YAP复杂网络调控机制，重点探索化合物或抑制剂激活Hippo和mTOR干扰该调控网络，或干扰IRF3/YAP，对肠上皮化生/上皮内瘤变的作用，为治疗胃癌前病变提供新的思路和理论依据。

Intestinal metaplasia and intraepithelial neoplasia are the precancerous lesions of gastric cancer. They are the key nodes of chronic gastritis transforming to gastric cancer or be cured, but the specific mechanism is unknown. Previous studies of our research group revealed that inflammatory factor TNF-α can directly or through NF-κB activate YAP to promote the inflammatory malignant transformation of gastric mucosa. Interfering YAP can significantly inhibit this event, suggesting that YAP plays an important role in gastric carcinogenesis. IRF3 is the positive regulator of YAP, and the target inhibition of IRF3/YAP can apparently inhibit the growth of gastric transplant tumors. Therefore, it is speculated that inflammatory factors can promote the inflammatory malignant transformation of gastric mucosa by phosphorylating YAP and regulating IRF3/YAP by mTOR through autophagy. This study is to construct the gastric mucosa inflammatory malignant transformation cell lines and PDX mice models, using lentiviral interference and chromatin immunoprecipitation method, to clarify the complex network regulation mechanism among Hippo-YAP, mTOR-YAP and NF-κB/IRF3-YAP, at mRNA level, protein level and post-translational modification levels. Our research will focus on the activation of Hippo and mTOR by compounds and inhibitors to interfere this regulating network, or IRF3/YAP, to determine the effect on intestinal metaplasia / intraepithelial neoplasia, and provide new ideas and theoretical basis for the treatment of gastric precancerous lesions.