Exosomes介导LncRNA-LIN0047调控miR-124在血管内皮细胞恶性转化中机制研究

Stephen Paget's original "seed and soil" hypothesis that proposed the organ-preference patterns of tumor metastasis are the product of favorable interactions between metastatic tumor cells (the "seed") and their organ microenvironment (the "soil"). Accumulating evidence indicates that exosomes play important roles in cell malignant transformation. Exosomes transfer oncogenic proteins and nucleic acids to modulate the activity of recipient cells and play decisive roles in tumorigenesis, growth, progression. Exosomes can act on various recipient cells. The uptake of exosomes may induce a persistent and efficient modulation of recipient cells. Our recent study found that tumor Exosomes promote endothelial cell malignant transformation，we also found that LncRNA-LIN0047 increased expression in tumor exosomes, and LncRNA-LIN0047 expression was correlated with miR-124. Firstly, we simulated tumor microenvironment to focus on the role of exosomes mediating LncRNA-LIN0047 in vascular endothelial cells of malignant transformation; furtherly, we studied the mechanism of recruitment of downstream signaling molecules and pathways after exosomes induced vascular endothelial cells malignant transformation to elucidate the molecular mechanism of these biological effects; finally, we compared LncRNA-LIN0047 knockdown of Exosomes on tumor microenvironment and cell signaling molecules and related vascular endothelium by construction of a mouse model to explore the mechanisms of exosomes- LncRNA-LIN0047 in cells malignant transformation, which provide a new theoretical basis for the regulation of exosomes mediation cell malignant transformation in microenvironment and the application of clinical oncology.

“种子与土壤”假说认为癌细胞通过血液和淋巴转移到其他组织，诱导周围细胞恶性转化，越来越多的研究发现，Exosomes在细胞恶性转化中起重要的作用。我们近期研究发现，肿瘤Exosomes促进血管内皮细胞恶性转化；深入研究发现，Exosomes中LncRNA-LIN0047表达升高，并与miR-124表达相关。本项目模拟肿瘤微环境，研究Exosomes介导LncRNA-LIN0047在血管内皮细胞恶性转化中的作用；进一步研究Exosomes诱导血管内皮细胞恶性转化后下游信号分子的募集和传导途径，阐明发生这些生物学效应的分子机制；最后构建小鼠模型，通过比较LncRNA-LIN0047 knockdown的Exosomes对肿瘤微环境及血管内皮细胞和信号分子的影响，探讨Exosomes-LncRNA-LIN0047在细胞恶性转化中的机制，为Exosomes调节血管内皮细胞恶性转化提供新的理论基础。

“种子与土壤”假说认为癌细胞通过血液和淋巴转移到其他组织，诱导周围细胞恶性转化，越来越多的研究发现，Exosomes在细胞恶性转化中起重要的作用。大量临床研究表明，乳腺癌转移的部位主要有肝、肺、骨骼和脑部，其中肺部为乳腺癌常见的转移部位。本项目我们首先围绕乳腺癌细胞分泌的外泌体来处理肿瘤微环境中血管内皮细胞、肺成纤维细胞以及肿瘤细胞。通过细胞增殖实验，细胞划痕实验以及Tranwell实验检测外泌体刺激的受体细胞恶性转化的差异；高通量RNA芯片测序检测受体细胞转录组表达；生物信息学以及RT-qPCR筛选验证相关LncRNAs的表达变化；通过构建LncRNAs相关表达载体在细胞内进一步研究其作用机制。结果表明乳腺癌来源的外泌体诱导肿瘤微环境中肿瘤细胞、血管内皮细胞、成纤维细胞增殖、侵袭等；其次我们进一步通过高通量芯片RNA-sequencing检测受体细胞转录组的表达变化，并结合生物信息学分析与细胞内实验验证，结果表明乳腺癌细胞外泌体刺激受体细胞后，多个LncRNAs发生显著上调或下降。最后通过构建动物实验模型，深入研究其机制表明，乳腺癌细胞外泌体通过介导LncRNA调控miR-124重塑乳腺癌肺部预转移微环境重塑。本课题的科学意义在于，我们揭示了乳腺癌细胞通过分泌外泌体介导的LncRNAs来重塑乳腺癌肺部预转移微环境。鉴于外泌体可以作为无创性分子诊断指标，本课题研究结果对于进一步研究外泌体在乳腺癌液态活检中临床应用奠定了理论基础与实验依据。项目相关研究成果共发表学术论文7篇，专利授权1项，获江苏省卫健委医学新技术引进奖二等奖1项。