循环非编码RNA调控肿瘤浸润免疫细胞NF-κB/STAT3网络在肝癌复发与转移中机制及诊断价值研究

Recently, plenty of studies indicated that chronic hepatitis is seriously related to development, metastasis and reoccurrence of liver cancer, some inflammation related protein factors have officially became circulating bio-marker in the diagnosis and treatment of liver cancer, whereas little is known about their up-stream regulating Non-coding RNA (ncRNA) . The present project will focus on ncRNA produced and secreted from immunocytes. High-throughput chipsets screening was applied to establish the metastasis and reoccurance associated database in liver cancer. An ncRNA/NF-κB/STAT3 signaling network was discovered by using bioinformatics analysis. Verification and correlation tests on some key molecules within this network will be carried out based on amount of clinical samplers, moreover, the impact of ncRNA abnormality on the malfunction of tumor infiltrating immunocytes will be further evaluated. The role of abnormal activation of ncRNA/NF-κB/STAT3 signaling network in tumor infiltrating immunocytes on development, metastasis and reoccurrence of liver cancer will be explored by a serial of experiments including in vitro co-culture of inflammatory and liver cancer cells, in vivo tumor growth, metastasis and chemical reagent induced mice hepatocarcinogenesis models. Furthermore, the diagnostic value of single and multiple joint key molecules within this signaling network in diagnosis of liver cancer will be seriously evaluated. The outcomes of this project will lay the theoretical foundation for a better interpretation of inflammation related hepatocarcinogenesis and find more sensitive circulating biomarkers in liver cancer patient.

近期研究表明慢性炎症与肝癌的发生、转移及复发密切相关，与慢性炎症相关因子已作为循环生物标志物应用于肝癌的诊治，但对于其上游具有调控效应的非编码RNA（ncRNA）认识较少。本研究聚焦于肝癌组织中浸润性免疫细胞产生并分泌的ncRNA，通过高通量芯片技术，构建肝癌转移与复发相关ncRNA数据库；结合生物信息学方法筛选得到与肝癌密切相关的ncRNA/NF-κB/STAT3信号网络，并利用大规模临床标本验证其中关键分子的表达，确定其表达相关性, 研究ncRNA表达改变对免疫细胞功能造成的影响；通过体外炎、癌细胞共培养，体内肿瘤生长、转移模型及化学试剂诱导的小鼠自发肝癌模型，共同分析该信号网络在免疫细胞中异常激活时促瘤促转移效应及机制，并研究该网络中关键分子单个或多个联合作为肝癌发生、转移及复发循环生物标志物的价值。本课题旨在从炎症角度更好的诠释肝癌发生，为发现更为敏感循环生物标志物奠定理论基础。

近期研究表明慢性炎症与肝癌的发生、转移及复发密切相关，与慢性炎症相关因子已作为循环生物标志物应用于肝癌的诊治，但对于其上游具有调控效应的非编码RNA（ncRNA）认识较少。本研究聚焦于肝癌组织中浸润性免疫细胞产生并分泌的ncRNA，通过高通量芯片技术，构建肝癌转移与复发相关ncRNA数据库；结合生物信息学方法筛选得到与肝癌密切相关的ncRNA/NF-κB/STAT3信号网络，并利用大规模临床标本验证其中关键分子的表达，确定其表达相关性, 研究ncRNA表达改变对免疫细胞功能造成的影响；通过体外炎、癌细胞共培养，体内肿瘤生长、转移模型及化学试剂诱导的小鼠自发肝癌模型，共同分析该信号网络在免疫细胞中异常激活时促瘤促转移效应及机制，并研究该网络中关键分子单个或多个联合作为肝癌发生、转移及复发循环生物标志物的价值。截至目前，我们已取得了许多重要成果：(1)长链非编码RNA Lnc-EGFR介导肿瘤浸润性免疫细胞对肝癌复发转移的研究；(2)CD97依赖G蛋白偶联受体经典信号通路促进肝细胞肝癌转移的机制研究；(3)TOX 在肝癌特异性CD8+ T 细胞耗竭中的调控作用及机制研究；(4)黄嘌呤氧化酶(XOR)在肝癌干细胞中的功能及其相关机制研究，等等。初步探究鉴定出多个参与调控肝癌发生、发展及转移相关的重要分子。对进一步开发肝癌早期诊断标志物，预后监测指标以及潜在治疗靶标具有重要的指导意义。