缺氧条件下miR-210-5p调控SPON2介导肝癌TAM细胞M2型极化参与耐药的机制研究

The anti-angiogenic drugs represented by sorafenib over the years have always been the first-line treatment of advanced liver cancer, but the drug resistance problem has always been a "bottleneck" in curative effect. Our previous study found that miR-210-5p is highly expressed in hepatocellular carcinoma and is closely related to sorafenib resistance. Combined with domestic and foreign research reports, we speculated that miR-210-5p might target to SPON2 to the polarization of M2-TAM in the induction of sorafenib resistance in HCC. Therefore, the mechanism of miR-210-5p targeted SPON2-mediated polarization of M2-TAM cell in hepatocellular carcinoma will be assessed and studied. The bioinformatics analysis method and IPA interaction network were used to screen and validate the polarization of M2-TAM cells involved in sorafenib resistance signaling pathway by combining the regression analysis of animal experiments and clinical samples. In all, it’s important to perform this study, thus providing a new strategy for intervention or modification of M2-polarization of TAM in hepatocellular carcinoma efficacy of anti-angiogenic drugs in the treatment of liver cancer.

多年来索拉非尼始终是晚期肝癌一线治疗药物，但其耐药问题一直是限制疗效的 “瓶颈”。我们前期研究发现miR-210-5p在肝癌中高表达并与索拉非尼耐药和乏氧调节密切相关，SPON2是miR-210-5p负调控的靶基因。结合国内外研究报道，推测miR-210-5p调控靶基因SPON2可能介导肝癌M2型肿瘤相关巨噬细胞（M2-TAM）极化参与诱导索拉非尼耐药。因此，本课题拟从分子、细胞、组织及实验动物水平，探索缺氧条件下miR-210-5p调控SPON2介导肝癌M2-TAM极化的分子机制；通过生物信息学方法和IPA互作网络，结合动物实验与临床样本回归性分析，筛选并验证M2-TAM极化参与索拉非尼耐药的信号通路，旨在阐明miR-210-5p调控SPON2介导肝癌M2-TAM极化参与索拉非尼耐药的作用机制，有望为干预或改造M2-TAM极化提供新的策略，为进一步提高肝癌的疗效提供潜在分子靶点。

本项目以索拉菲尼耐药与缺氧微环境中的TAM极化对肝癌的影响为切入点，深入探讨并明确了缺氧诱导的miR-210-5P对肝癌细胞索拉菲尼敏感性及其恶性生物学行为的影响，SPON2与肝癌发生发展中的相关性及其介导TAM极化对肝癌恶性表型的影响及分子机制，取得多项有意义的结果：（1）缺氧能诱导miR-210-5p上调促进肝癌细胞迁移和EMT改变，抑制miR-210-5p可增加肝癌细胞对索拉非尼的敏感性。（2）缺氧可以上调肝癌细胞SPON2的表达。（3）SPON2的表达与肝癌预后正相关，干扰SPON2表达可以诱导肝癌细胞发生周期阻滞，诱导细胞凋亡。（4）M1/M2-TAM分布差异与患者的预后及临床病理特征密切相关，相对于癌旁组织，M2-TAM在癌组织中表达水平越高，患者预后越差。（5）SPON2介导M1/M2-TAM极化对M1/M2-TAM分布差异造成影响。研究结果说明肿瘤治疗成败与肿瘤微环境状态密切相关，通过干预SPON2改变M1/M2-TAM在癌组织中的分布可能是提高肝癌治疗预后的有效途径。