肿瘤相关性中性粒细胞和巨噬细胞相互作用通过激活肝内胆管细胞癌STAT3信号通路促进肿瘤侵袭转移的机制研究

Tumor-associated neutrophils (TANs) and macrophages (TAMs) can influence cancer growth and metastasis, but their association and the mechanism on intrahepatic cholangiocarcinoma (ICC) progression remains unclear. We have found that TANs and TAMs after inter-coculture can enhance the proliferation and invasion ability of ICC cells in vitro. PCR array results showed that TANs abundantly produced OSM, while IL-11 secretion was significantly increased by TAMs, both of which activated STAT3 signal in ICC cells. In this study, we firstly plan to identify by in vitro and in vivo study,OSM/IL-11/STAT3 which involved in the metastasis and recurrence of ICC and then explore the STAT3 signal pathway involved in epithelial-mesenchymal transition (EMT) which regulated by TANs and TAMs inter-coculture. We will perform by probing a human phospho-kinase array to identify the activated signaling pathways of TANs and TAMs after inter-culture. Finally, using tissue microarrays (TMAs) and immunohistochemical in ICC samples, we will determine the relationship between TANs and TAMs infiltration, and evaluate their prognostic significance. Through the above research, we will determine that TANs and TAMs interaction contributes to progression of ICC by activation of STAT3.

肿瘤相关性中性粒细胞（TANs）和巨噬细胞（TAMs）在肿瘤发生发展过程发挥重要作用。前期工作发现肝内胆管细胞癌（ICC）微环境中TANs和TAMs共培养促进ICC细胞增殖侵袭；TANs和TAMs共培养后分别高表达细胞因子OSM及IL-11，均激活ICC的STAT3。本课题拟首先探讨ICC微环境中TANs/TAMs分泌的OSM/IL-11以及ICC细胞的STAT3在肿瘤侵袭转移过程中的重要作用；进一步挖掘TANs和TAMs共培养后促进ICC侵袭转移及发生上皮-间充质转化的分子机制；明确TANs和TAMs共培养后上调表达OSM及IL-11的信号途径；最后在临床样本中观察TANs、TAMs、p-STAT3及下游信号分子的表达和定位，并对其预后价值进行评价。通过以上研究，明确ICC微环境中TANs和TAMs的相互作用促进肿瘤侵袭转移的作用机制，探讨以间质细胞作为潜在肿瘤分子靶点的可行性。

本课题以肝内胆管细胞癌为研究对象，研究肿瘤微环境中炎症细胞之间的相互调控，通过分泌细胞因子OSM和IL-11激活STAT3信号通路促进肿瘤转移复发的作用机制，深入探讨以间质细胞作为潜在肿瘤分子靶点的可行性，有助于开阔肿瘤复发转移治疗的新思路。.本课题通过体外细胞增殖、迁移、侵袭实验和体内小鼠成瘤试验，首先明确了TANs和TAMs分泌的OSM/IL-11以及肝内胆管细胞癌的STAT3，在TANs和TAMs共同培养后，促进ICC侵袭转移的重要作用；然后观察TANs和TAMs共同培养对ICC细胞上皮-间充质转化（EMT）标志物表达及亚细胞定位的影响，进一步明确了TANs和TAMs共同培养后诱导ICC发生EMT促侵袭转移的信号调控网络；同时，应用磷酸化蛋白芯片联合western blot、ELISA检测，阐明TANs和TAMs共同培养后激活TANs和TAMs的信号分子，明确共培养后上调表达OSM及IL-11的信号途径；最后，在ICC病人制作的组织芯片中利用免疫组织化学和免疫荧光染色，准确观察TANs、TAMs、p-STAT3及下游信号分子的表达和定位，并对其预后价值进行评价。该课题阐明肿瘤相关性中性粒细胞和巨噬细胞存在相互作用，并分泌细胞因子OSM 和IL-11 激活肝内胆管细胞癌的STAT3 信号通路，进而促进肿瘤转移复发，为探讨预防肿瘤转移复发中提供了新的干预靶点。