锦灯笼酸浆苦素调控LIX1L重塑肿瘤微环境的抗肝癌机制研究

肿瘤微环境所致的肝癌复发和治疗耐受是临床治疗过程中亟待解决的瓶颈问题。干预和改善微环境是中医药治疗疾病的优势所在。趋化因子参与构建炎性和免疫抑制微环境促进肝癌增殖和转移。前期研究发现LIX1L上调趋化因子CCL20表达促进肝癌细胞增殖和转移，且与肝癌病人预后差密切相关，而中药锦灯笼特征成分酸浆苦素类能下调LIX1L蛋白抑制CCL20表达，提示其抗肝癌作用与调控肿瘤微环境有关。本项目拟通过Biacore、质谱、免疫共沉淀等方法确定酸浆苦素下调LIX1L的分子机制；建立化学诱导的肝癌模型、肝癌细胞皮下和原位移植瘤模型，确证酸浆苦素通过LIX1L-CCL20信号轴重塑肝癌微环境抑制肝癌进展的机制；利用蛋白组学、RNA-seq及生物信息学分析酸浆苦素对炎症和免疫信号通路的影响，最终阐明酸浆苦素的作用靶点、分子机制及信号调控网络，为锦灯笼酸浆苦素应用于肝癌治疗及克服临床治疗耐受提供新思路和理论依据。

Tumor microenvironment induced liver tumorigenesis, relapse, and resistance are the bottlenecks to be solved in clinic. Intervention and improvement of the microenvironment are the advantages of traditional Chinese medicine for treating diseases. Chemokines-mediated inflammatory and immunosuppressive microenvironment promotes liver cancer cell proliferation and metastasis. Previously, we found LIX1L protein promoted hepatoma cell proliferation and metastasis via up-regulation of chemokine CCL20, which was closely related to the poor prognosis of liver cancer patients. However, physalin, the characteristic and principal bioactive substances of Jin-Deng-Long, could down-regulate LIX1L protein expression and inhibit CCL20 expression, suggesting its potential effect on tumor microenvironment remodeling. Based on these data, this project intends to determine the molecular mechanism of physalin induced down-regulation of LIX1L by Biacore, mass spectrometry and co-immunoprecipitation; establishing chemical induced HCC model, subcutaneous and orthotopic HCC model, to confirm that physalin suppresses HCC progress by remodeling tumor microenvironment via LIX1L-CCL20 axis; employing proteomics, RNA-seq, and bioinformatics analysis to investigate the effect of physalin on inflammatory and immune signaling pathways, finally uncover the target, molecular mechanisms, and signaling network. This study provides new insights and theoretical basis for the application of physalin in the treatment of liver cancer and to overcome clinical treatment tolerance.