巨噬细胞内HSP27表达调控其亚型转化并影响肝癌侵袭转移的作用机制

项目名称： 巨噬细胞内HSP27表达调控其亚型转化并影响肝癌侵袭转移的作用机制

项目编号： No.81472218

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 代智

作者单位： 复旦大学

项目金额： 85万元

中文摘要： 肿瘤相关巨噬细胞(TAMs)是肿瘤免疫微环境中的重要成分，在肿瘤转移中发挥重要作用。前期工作中,我们在发现肝癌组织中TAMs特异性标志物的基础上，率先揭示巨噬细胞内HSP27过表达可显著增加肝癌细胞运动侵袭能力,且与巨噬细胞向M2亚型转化和肝癌细胞上皮-间质转化（EMT）密切相关。本课题模拟人体肝癌免疫微环境建立体内和体外模型，通过干预巨噬细胞中HSP27表达，观察巨噬细胞生长凋亡、运动和亚型转化的变化，并明确HSP27在巨噬细胞诱导肝癌EMT和侵袭转移中的作用；应用抗体芯片筛选巨噬细胞中HSP27表达调控的与其亚型转化和肝癌发展相关的关键分子；应用共转染、免疫印迹和免疫共沉淀分析HSP27对关键分子表达的影响及二者相互作用关系，同时分析它们在肝癌组织中的表达及其与肝癌转移复发的关系，并进行体内、外功能分析，明确其在肝癌转移中的作用。通过上述研究，为探讨肝癌治疗中新的干预靶点提供实验依据。

中文关键词： C09_肝和肝内胆管肿瘤；热休克蛋白27；巨噬细胞；肿瘤转移

英文摘要： Tumor-associated macrophages (TAM) are a major component of the cancer-related microenvironment and play a central role in tumor metastasis. In our previous study, we found that the specific marker CD206 was found to identify TAMs in hepatocellular carcinoma (HCC) tissues. Furthermore, it was found that overexpression of HSP27 in macrophages could promote invasive ability of HCC cell lines in vitro; this effect may be associated with macrophage subsets transformation and the epithelial-mesenchymal transition (EMT) of HCC cells. This was a novel finding of the impact of HSP27 in macrophages on cancer progression. This study will establish in vivo and in vitro model of human HCC immune microenvironment.Using the model, we will investigate whether HSP27 is involved in regulation of macrophage suvival, migration and phenotype switch. In addition, through short hairpin RNA (shRNA) interference or up-regulation of HSP27 expression in macrophages, we will determine the role of HSP27 in macrophage-induced HCC invasion and the EMT process. Furthermore, we will use antibody arrays to screen HSP27-regulated key proteins involved in macrophage subsets transformation and HCC progression. Western blotting and co-immunoprecipitation will be used to analyze the interaction between HSP27 and the key proteins, and investigate the role of HSP27 expression on regulation of proteins. In addition, the relationship between the expression of these proteins and the invasive phenotype of HCC cells will be studied. We will determine whether these proteins could be important independent factors in determining clinical outcomes of HCC. Using specific RNA interference or up-regulation of these genes in HCC cells, we will assess their roles in tumor invasion and in vivo metastasis. This study will provide a biological foundation for the discovery of new potential therapeutic targets for HCC.

英文关键词： Hepatocellular carcinoma;Hsp27;Macrophage;Tumor metastasis