麦冬皂苷通过下调lnc-MALAT1抑制NSCLC血管生成的机制研究

项目名称： 麦冬皂苷通过下调lnc-MALAT1抑制NSCLC血管生成的机制研究

项目编号： No.81503374

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 陈美娟

作者单位： 南京中医药大学

项目金额： 18万元

中文摘要： 血管新生是NSCLC转移的关键环节，中医“养阴”疗法对于控制NSCLC转移具有显著的效果。麦冬是典型的养阴类中药，而麦冬皂苷是其重要的活性成分。前期研究表明，麦冬皂苷能显著抑制A549移植瘤裸鼠肿瘤血管生成，并抑制其EphA2/Akt信号通路。lnc-MALAT1是与NSCLC侵袭转移密切相关的长链RNA，生物信息学分析发现其启动子区存在多个EphA2/Akt通路下游转录因子HIF-1的结合位点。基于以上研究发现，本项目拟通过CAM接种实验和matrigel plug assay研究麦冬皂苷抑制NSCLC血管生成的表型；通过luciferase assay、EMSA和ChIP等实验，确定麦冬皂苷通过HIF-1调控lnc-MALAT1转录水平，进一步影响NSCLC血管生成的作用，以初步阐明麦冬皂苷抗NSCLC的分子机制，并为“养阴”疗法用于临床抑制NSCLC转移提供一定的科学依据。

中文关键词： 非小细胞肺癌；血管生成；麦冬皂苷；EphA2/Akt/HIF-1α；长链非编码RNA-MALAT1

英文摘要： Angiogenesis is critical during the metastasis of NSCLC. “Nourish yin” therapy of traditional Chinese medicine is effective on inhibiting metastasis. Radix Ophiopogonis has significant effect of “nourish yin”, and ophiopogonin being the important active ingredient of it. Our previous study found that ophiopogonin inhibited tumor angiogenesis of A549 xenografts in nude mice and inhibited EphA2/Akt signaling pathway in NSCLC cell lines. Lnc-MALATA1is critical in regulating metastasis and migration of NSCLC, bioinformatics analysis found that transcription factor HIF-1α which locating downstream of EphA2/Akt pathway had several binding-site in the promoter region of lnc-MALATA1.Based on the preliminary research findings, this project is programmed to investigate the inhibition mechanism of ophiopogonin on angiogenesis in NSCLC. Phenotype of angiogenesis will be tested by chicken chorioallantoic membrane assay and matrigel plug assay. Regulation of HIF-1α on the transcription level of lnc-MALATA1 will be investigated by luciferase assay, EMSA and ChIP assay. It will be helpful for elucidating the molecular machanisms of ophiopogonin on the angiogenesis of NSCLC, and providing scientific basis for the use of “nourish yin” therapy in clinical treatment of NSCLC.

英文关键词： NSCLC;angiogenesis;ophiopogonin;EphA2/Akt/HIF-1α ; long non-coding RNA-MALAT1