长链非编码RNA LINC01296调控ASPP2表达促进肝细胞肝癌转移的机制研究

项目名称： 长链非编码RNA LINC01296调控ASPP2表达促进肝细胞肝癌转移的机制研究

项目编号： No.81502047

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： 黄明德

作者单位： 南京医科大学

项目金额： 17万元

中文摘要： 转移为肝细胞肝癌患者死亡的重要原因且其分子机制仍未完全阐明。表观遗传学的改变如lncRNA参与了肿瘤转移的过程。课题组发现LINC01296在肝癌中表达显著上调，其表达与肿瘤大小、分期及转移呈显著正相关；干扰LINC01296后肝癌细胞侵袭转移能力明显下降，并上调ASPP2表达；RIP实验证实LINC01296与PRC2结合，ChIP实验证实PRC2与ASPP2启动子区结合。生物信息学发现LINC01296启动子区有2个E2F1结合位点，干扰E2F1后其表达下调3.4倍。据此提出假说：肝癌中E2F1促进LINC01296表达，LINC01296通过绑定PRC2介导ASPP2启动子区H3K27三甲基化抑制其转录，促进肝癌转移。本课题将通过临床样本验证，运用RIP、ChIP、荧光素酶报告基因及动物模型等实验证实上述假设，丰富肝癌转移机制，为肝癌的治疗提供新的靶点。

中文关键词： 肝和肝内胆管肿瘤；长链非编码RNA；转移；LINC01296

英文摘要： Metastasis is the important reason for death in hepatocellular carcinoma(HCC) and the mechanism of which is still not well documented. Long noncoding RNA(lncRNA) might have critical functions in cancer metastasis. We applied bioinformatics analysis and qPCR to detect LINC01296 expression in tumoral tissues and paired nontumoral tissues from HCC patients. LINC01296 expression was up-regulated in HCC tissues and higher expression of LINC01296 was significantly correlated with tumor size, BCLC stage and metastasis. Moreover, we found that knockdown of LINC01296 could inhibit HCC cells migration and invasion in vitro. We also found that LINC01296 could bind with PRC2 and epigenetically repress ASPP2 transcription in HCC cells. Bioinformatics analysis showed that LINC01296 promoter region contained two E2F1 binding sites. LINC01296 was downregulated by 3.4 folds while inhibition of E2F1 in HCC cells. Accordingly, the hypothesis is proposed:E2F1 promotes LINC01296 expression, and LINC01296 represses ASPP2 expression by binding with PRC2 and recruiting it to ASPP2 promoter region. We will document this hypothesis by clinical samples, RIP, ChIP, luciferase reporter assay and animal models et.al methods. Our work will further the understanding about the molecular mechanisms of HCC metastasis and provide a new therapy target for HCC.

英文关键词： liver cancer; long noncoding RNA; metastasis;LINC01296