E2F1调控的长链非编码RNA LINC00668调节细胞周期促进胃癌增殖机制的初步研究

项目名称： E2F1调控的长链非编码RNA LINC00668调节细胞周期促进胃癌增殖机制的初步研究

项目编号： No.81502071

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： 殷丹丹

作者单位： 东南大学

项目金额： 18万元

中文摘要： LncRNAs普遍参与肿瘤细胞增殖、凋亡、侵袭转移的调控过程。生物信息学分析发现在胃癌中lncRNA LINC00668明显高表达，并在组织中得到验证。同时发现E2F1能够诱导LINC00668表达。细胞实验证实LINC00668能够明显影响胃癌细胞的增殖和周期功能，并影响CKIs和CDK6表达。RIP实验证明LINC00668在细胞核中能够和PRC2绑定，序列分析发现LINC00668在细胞质中可能能够“吸附”miR-449a而调控CDK6表达。据此提出以下假设：在胃癌中，高表达的E2F1诱导LINC00668高表达，在细胞核表观抑制CKIs转录，在细胞质中发挥ceRNA活性，竞争性吸附miR-449a，释放CDK6表达，从而加速胃癌细胞周期进程，引起胃癌细胞恶性增殖。本课题组将扩大临床样本量，并用相关技术证实上述假设，丰富胃癌发生、发展的分子机制研究，为临床胃癌诊断、治疗提供理论依据。

中文关键词： E2F1；LINC00668；胃肿瘤；细胞周期；细胞增殖

英文摘要： Recent studies indicated that lncRNAs involve in regulating tumor cells proliferation, apoptosis, invasion and metastasis. By utilizing publicly available lncRNAs expression profiling data of gastric cancer and integrating analyses, we screened out LINC00668, whose expression is significantly increased in gastric cancer. Moreover, E2F1 could induce the expression of LINC00668. Further experiments confirmed LINC00668 could significantly affect gastric cancer cell proliferation, cell cycle and affect CKIs and CDK6 expression. RIP assay proved that LINC00668 could bind to PRC2 in the nucleus, and sequence analysis revealed LINC00668 may be able to “sponge miR-449a expression and regulation of CDK6 in the cytoplasm. Accordingly, the hypothesis is proposed: overexpression of E2F1 induces the expression of LINC00668. In the nucleus, LINC00668 could epigenetically silence expression of CKIs. In the cytoplasm, LINC00668 could serve as a “ceRNA”, competitive adsorption of miR-449a, releasing CDK6 expression, thus speeding up gastric cancer cell cycle progression, causing malignant proliferation of gastric cancer cells.We will documente this hypothesis by clinical samples, nude mice, ChIP, RIP, Luciferase reporter and RNA-pull down et.al. Moreover, our work will further the understanding about the molecular mechanisms of development of gastric cancer and provide a new basis for gastric cancer clinical individual therapy and molecular targeted therapy.

英文关键词： E2F1;LINC00668;gastric cancer ;cell cycle;cell proliferation