基于网络互作研究雷丸蛋白pPeOp调控Cdc42/JAK/STAT3信号通路抑胃癌作用机制

胃癌是临床常见恶性肿瘤，多以手术切除辅以放、化疗治疗，但疗效不显著，若开发多靶点、多途径抗肿瘤药物，将显著提高疗效。本项目前期研究表明，抗肿瘤中药雷丸所含pPeOp蛋白负调控JAK/STAT3通路抗胃癌时，可显著抑制Cdc42的表达与活性。鉴于Cdc42是STAT3磷酸化的另一重要调控因子，我们推测pPeOp蛋白的抗胃癌作用可能与Cdc42/JAK/STAT3通路的网络调控有关。因此，本申请项目拟采用慢病毒转染胃癌细胞，构建Cdc42和STAT3沉默细胞并制备荷瘤裸鼠模型。IP和ChIP-qPCR分析pPeOp蛋白干预后，Cdc42和STAT3与胞内蛋白及靶基因互作，网络药理学方法分析mRNA、miRNA和蛋白互作效应，确定pPeOp蛋白对Cdc42/JAK/STAT3通路的网络调控作用，明确关键调控靶点，阐明pPeOp蛋白的抗胃癌作用与Cdc42/JAK/STAT3通路网络调控的相关性。

Gastric cancer is one of the most common clinical malignant tumor, which be treated by surgery auxiliary with chemotherapy and radiotherapy. However, the efficacy of such treatment is not ideal as expectable. Hence, it`s very important to develop multi-target and multi-pathway anti-cancer drugs to significantly improve the efficacy and survival rate of patients. In our preliminary study, the results suggested that pPeOp protein from Omphalia lapidescen has significantly down-regulate the JAK/STAT3 signaling pathway, meanwhile down-regulate the expression and activity of Cdc42 protein. Since Cdc42 is a key regulatory factor in Cdc42 phosphorylation, we speculate that the anti-tumor effect of pPeOp should be related with Cdc42/JAK/STAT3 signal pathway. Therefore, the present study intends to establish Cdc42 and STAT3 gene silencing cell models by lentiviral transfection, and further to establish the naked mice model with the transfected gastric cancer cells. Meanwhile, making the wild type cells as the control group, we try to detect the variation of Cdc42/JAK/STAT3 signaling pathway in gastric cancer cells and tumor body after the pPe0p protein treatment. Furthermore, we tend to analyze the mRNA interaction, miRNA interaction and protein interaction by network pharmacology and bioinformatics, and to analyze the effect of pPeOp protein on Cdc42/JAK/STAT3 network. Then, the key regulatory nodes of Cdc42/JAK/STAT3 signaling pathway would be identified and validated. Finally, we propose to clarify the anti-gastric cancer target network of pPe0p protein with in vitro and in vivo experimental data.