一个功能未知的长链非编码RNA——AC093850.2调控肺腺癌侵袭转移的分子机制研究

项目名称： 一个功能未知的长链非编码RNA——AC093850.2调控肺腺癌侵袭转移的分子机制研究

项目编号： No.81472200

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 尹荣

作者单位： 南京医科大学

项目金额： 78万元

中文摘要： AC093850.2（简称AC）是我们前期应用芯片及二代测序技术筛选获得的一条特异高表达于肺腺癌、功能未知的长链非编码RNA。已发现AC在肺腺癌中高表达程度与淋巴结转移程度正相关；沉默AC可显著抑制肺腺癌细胞侵袭能力；生物信息学分析和实验提示AC与染色质修饰复合物PRC2相互结合；沉默AC后PRC2靶基因E-cad显著上调；临床标本中AC与E-cad表达水平高度负相关。因此我们提出AC招募PRC2调控E-cad等肺癌转移相关基因促进肺腺癌侵袭转移的科学假说。本研究拟采用过表达/沉默策略，结合体内体外实验，论证AC在肺腺癌侵袭转移中的作用；应用RIP、ChIP、ChIRP等技术及拯救实验，深入阐明AC招募PRC2调控E-cad的分子机制；最后大样本病例评价AC、PRC2及E-cad表达在肺腺癌诊疗中的临床价值。有关AC促进肺腺癌侵袭转移的作用和机制未见报道，本研究可望为肺腺癌防治提供新靶标。

中文关键词： C05_气管；支气管；肺肿瘤；长链非编码RNA；AC093850.2；转移；机制

英文摘要： By microarray and transcriptome sequencing, we have identified an uncharacterized long non-coding RNA (lncRNA) AC093850.2,which is highly expressed in lung adenocarcinoma (LAC). Quantitative RT-PCR demonstrated that AC093850.2 was significantly over-expressed in LAC tissues compared with adjacent normal lung tissue and positively correlated with lymph node metastasis. AC093850.2 was also specifically highly expressed in LAC cell lines but not in other epithelium derived cancer cells or in lung saqumaous cell carcinoma. Silencing AC093850.2 by siRNA significantly suppressed the invasion and migration ability of LAC cell lines. Moreover, bioinformatic analysis and RIP-PCR experiment revealed that AC093850.2 could bind with an important chromosome remodeling complex, polycomb repressive complex 2 (PRC2), indicating the potential of epigenetic regulation. Microarray and in vitro validation suggested that silencing AC093850.2 significantly up-regulated the expression of a lung cancer metastasis suppressor gene , E-cadherin, which is also a target gene of PRC2. Based on above results, we therefore hypothesized that AC093850.2 might suppress the metastasis of LAC by recuiting PRC2 and subsequently decreasing the expression of E-cadherin. In this proposal, firstly, we plan to clarify the functional role of AC093850.2 in invasion and metastasis of LAC. To achieve this goal, we design to over-express and knock-down AC093850.2 both in vivo (in both passitive and initiative metastatic NOD/SCID mice model) and in vitro (in two separated LAC cell lines) and evaluate the impact of AC093850.2 on migration and invasion. Secondly, we aim to explore how AC093850.2 exerts its regulatory function. Highlighted by the relationship between E-cadherin gene and AC093850.2, we design to investigate the mutual interaction between E-cadherin and AC093850.2 via RIP, ChIP, ChIRP, and rescue experiments.Finally, we design to evaluate the expression level of AC093850.2, EZH2, and E-cadherin in a high through-put human LAC tissue microarray, as well as their correlation with clinical profiles. Our study is the original source of innovation and can provide new potential biomarkers for the prevention and treatment of LAC.

英文关键词： lung cancer;lung non-coding RNA;AC093850.2;metastasis;mechanism