aPKC-ι/TNFβ诱导TAMs活化并调控胆管癌细胞极化的研究

癌细胞极化状态的改变是肿瘤侵袭转移的核心环节，肿瘤免疫微环境的改变也是影响癌细胞侵袭转移的重要因素。申请者前期研究发现，aPKC-ι（调节细胞极化的关键蛋白）高表达的胆管癌细胞可诱导TAMs（肿瘤免疫微环境中最丰富的浸润性免疫细胞）活化，并初步发现，高表达aPKC-ι的胆管癌细胞培养液上清中TNFβ分子水平明显上调。申请者据此推测：aPKC-ι/TNFβ通路可通过调控胆管癌细胞极化影响TAMs活化，并介导癌细胞侵袭转移。本项目拟在胆管癌细胞体外侵袭转移的极化运动实验模型基础上，观察aPKC-ι高表达或失活对胆管癌细胞极化状态的调控作用，及对TAMs功能的影响；探讨aPKC-ι调控TNFβ表达的分子机制；通过干扰aPKC-ι/TNFβ通路，观察对胆管癌细胞侵袭转移的影响。本项目可望从新的角度阐明胆管癌细胞侵袭转移的分子机制，并为探索胆管癌治疗的新策略提供依据。

Polarization motion is a core aspect of the invasion and metastasis in tumor initiation stage, meanwhile tumor immune microenvironment is an important factor affecting the invasion and metastasis of cancer cells. In the previous studies, the applicant firstly found that as a key polarization regulatory protein, the cholangiocarcinoma cells with highly expressed aPKC-ι could induce the activation of TAMs which was the most abundant infiltrating immune cells in tumor immune microenvironment. The applicant also found that the level of TNFβ in culture supernatant of cholangiocarcinoma cells with highly expressed aPKC-ι was upregulated. Based on the above statements, the applicant infer that aPKC-ι/TNFβ could regulate the cell polarization to infulence the activation of TAMs, then to induce the invasion and metastasis in cholangiocarcinoma. This project intends to establish the polarization movement experimental model of the invasion and metastasis of cholangiocarcinoma cells in vitro, studies the regulation of the cell polarization state of cholangiocarcinoma cells and its effects on TAMs cell function when aPKC-ι overexpressed or inactived, investigates the molecular mechanisms of aPKC-ι regulating TNFβ, and studies the effects on invasion and metastasis by interfering aPKC-ι/ TNFβ signaling pathway. The project is expected to clarify the molecular mechanisms on invasion and metastasis of cholangiocarcinoma cells from a new perspective and it suggests us a new therapeutic approach for the treatment of cholangiocarcinoma.