HOTAIR/miR-326/SP1调控通路对非小细胞肺癌增殖、迁移和侵袭能力的影响及作用机制

项目名称： HOTAIR/miR-326/SP1调控通路对非小细胞肺癌增殖、迁移和侵袭能力的影响及作用机制

项目编号： No.81502513

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： 王蓉

作者单位： 南京医科大学

项目金额： 18万元

中文摘要： microRNA在多种恶性肿瘤的发生发展过程中发挥重要的调控作用，但目前对一些microRNA的作用机制还有待进一步探讨。我们预实验结果提示miR-326可能是肺癌的抑癌基因，其在肺癌组织中表达下调，伴随HOTAIR和SP1表达上调，生物信息学预测HOTAIR与SP1分别有miR-326上游和下游结合位点。故提出假说：HOTAIR能够从上游调控miR-326的表达，同时miR-326可以调控其下游靶基因SP1的表达，从而构成HOTAIR/miR-326/SP1调控通路，影响肺癌细胞增殖、迁移和侵袭。为验证这一假说，我们将利用肺癌细胞系、裸鼠成瘤模型及临床组织样本，采用real time PCR、Western blot、腺病毒载体转染、RNA干扰、荧光素酶报告及CHIP等实验手段，从分子、细胞、组织和动物整体水平阐明该通路的调控功能和作用机制，为肺癌增殖和转移的发生机制奠定新的理论基础。

中文关键词： C05_气管；支气管；肺肿瘤；microRNA-326；长链非编码RNA；HOTAIR；SP1；增殖和侵袭

英文摘要： microRNA plays a crucial role in development and progression of multiple malignancies. However, the mechanisms of microRNA in cancer are not totally clear. Our previous study showed miR-326 may function as a tumor suppressor in lung cancer. MiR-326 downregulated in lung cancer specimens with lymph node metastasis. Conversely, lncRNA HOTAIR and SP1 upregulated in metastatic lung cancer specimens. In addition, bioinformatics analysis showed miR-326 and SP1 binding sites were found in HOTAIR sequence and in the promoter of HOTAIR, respectively. Therefore, we hypothesize that tumor related lncRNA HOTAIR is a upstream regulator of miR-326, and that SP1 is a downstream target of miR-326 and regulated by miR-326. HOTAIR, miR-326 and SP1 communicates with each other and comprise a miR-326-based regulation network, which is crucial for proliferation, metastasis and invasion of lung cancer. To verify our hypothesis, lung cancer cell lines A549 and H838, nude mince with lung cancer xenograft and lung cancer tissue samples are used. Real time PCR, western blot, adenovirus, RNA interference, luciferase assays and chromatin immunoprecipitation assays (CHIP) are employed to investigate molecular mechanisms of HOTAIR/miR-326/SP1 regulation pathway in lung cancer. Our project will provide new information for lung cancer development and progression, and help establish novel strategy for lung cancer diagnosis and therapy.

英文关键词： non-small cell lung cancer(NSCLC);microRNA-326; lncRNA HOTAIR;SP1;proliferation and invasion