一个功能未知的长链非编码RNA PIK3CD-AS2调控非小细胞肺癌侵袭转移的机制研究

项目名称： 一个功能未知的长链非编码RNA PIK3CD-AS2调控非小细胞肺癌侵袭转移的机制研究

项目编号： No.81501977

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： 王洁

作者单位： 南京医科大学

项目金额： 18万元

中文摘要： PIK3CD-AS2（简称PI）是前期应用芯片和TCGA大数据挖掘筛选到的一条在人非小细胞肺癌（简称肺癌）组织中差异高表达、功能未知的长非编码RNA。前期发现PI在肺癌组织中显著高表达，与淋巴结转移程度正相关，且表达越高预后越差；沉默PI后可显著抑制肺癌细胞株侵袭转移能力；进一步预实验和生物信息学分析提示癌基因ECT2是PI潜在下游靶点，PI可能通过结合甲基转移酶核心亚基WDR5表观上调ECT2表达、促进肺癌侵袭转移。故我们提出PI招募WDR5表观上调ECT2促进肺癌侵袭转移的科学假说。本研究拟临床评价PI与肺癌侵袭转移的关系；结合过表达/沉默策略，在细胞和动物水平共同评价PI调控肺癌侵袭转移的生物学功能；设计拯救实验，应用ChIP、RIP等技术，阐明PI通过表观调控ECT2表达、促进肺癌侵袭转移的分子机制。有关PI促进肺癌侵袭转移的功能和机制尚未见报道，本研究将可为肺癌防治提供新靶标。

中文关键词： 肺肿瘤；长链非编码RNA；侵袭转移；机制

英文摘要： Using microarray analysis and big data from TCGA project, we identified a novel long noncoding RNA (lncRNA), PIK3CD-AS2 (PI), with unknown function that is highly expressed in non-small cell lung cancer (NSCLC) tissues. Our previous work has demonstrated that PI is significantly overexpressed in in NSCLC tissues, positively correlated with lymph node metastasis, and negatively correlated with prognosis. Knockdown of PI significantly inhibited migration and invasion of NSCLC cells. By mining of bid data and experiment, we found that ECT2 could be the downstream target of PI and we hypothesized that PI could promote the invasion and metastasis of NSCLC by epigenetic activation of ECT2 via binding to WDR5, a core subunit of histone 3 lysine 4 methyltransferase. Here, we plan to validate our hypothesis by assess the relationship between PI expression level and NSCLC metastasis; investigate the influence of PI on invasion ability of NSCLC both in vivo and in vitro in over-expression/knockdown manner. The rescue experiment along with chromatin immunoprecipitation, RNA immunoprecipitation, and RNA pull down assays were designed to underlying molecular mechanism by which PI epigenetically regulates ECT2 expression and promotes NSCLC invasion. There is no report about PI function so far and our study offers a new therapeutic target for the diagnosis and treatment of NSCLC.

英文关键词： lung cancer;long non-coding RNA;metastasis;mechanism