长链非编码RNA URHC调控ZAK介导肝癌生长分子机制的研究

The relationship between LncRNA (long non-coding RNA) and cancer has become a heated topic in recent years. We recently identified a new LncRNA in HCC (Hepatocellular Carcinoma), URHC (Up-regulated in HepatoCellular Carcinoma), which was closely related to the growth of HCC. We reported that URHC promoted the growth of HCC via a ZAK-dependent way (Int J Biol Sci. 2014.), but the mechanism of which regulates the expression of ZAK remains unknown. Based on previous reports and our studies, we propose that URHC promote the growth of HCC via regulating the expression of ZAK in cis. By RNA pull down, ChIRP, RIP, Mass spectrometry and so on, we will reveal the cis-regulatory effect of URHC on the down-regulated expression of ZAK from cell and molecular level, and also confirm the key downstream effectors of URHC-ZAK signal; further explore the relation between URHC-ZAK signal and tumor prognosis, the clinical and pathological phenotypes; By in-vivo imaging technology, finally explore the possibility and feasibility of magnetic nanoparticles encapsulating URHC siRNA (Nanomedicine. 2013.), inhibiting the growth of HCC. This project will systematically illuminate the molecular mechanism by which URHC regulates the expression of ZAK to promote the growth of HCC, and provide effective biomarkers and potential targets for the clinical diagnosis, prognosis and prevention of HCC.

长链非编码RNA是近年肿瘤研究热点之一。近来，我们在肝癌中鉴定并命名一条与肝癌生长密切相关的长链非编码RNA URHC，证实URHC通过ZAK依赖性途径促进肝癌生长（Int J Biol Sci. 2014），但其调控ZAK表达的机制不明。结合文献报道和我们的研究结果，我们提出“URHC顺式调控ZAK介导肝癌细胞生长”的假说。本项目拟从细胞分子水平，利用RNA pull down、ChIRP、RIP及质谱等揭示URHC对ZAK的顺式调控作用，明确URHC-ZAK下游关键效应分子；从肝癌组织水平，评价URHC-ZAK通路在肝癌诊断和预后判定中的临床价值；从动物水平，利用活体成像探讨磁性纳米介导的URHC siRNA(Nanomedicine. 2013)抑制肝癌生长的可能性和可行性；系统阐明URHC调控ZAK介导肝癌细胞生长的分子机制，为临床肝癌诊断、预后及防控提供有效生物学标志和潜在靶点。

长链非编码RNA(long noncoding RNA, lncRNA)是肿瘤研究热点之一，研究表明其在肝细胞癌（hepatocellular carcinoma, HCC）进程中起重要作用。前期我们在肝癌鉴定并命名一条与肝癌生长密切相关的LncRNA-URHC (LncRNA up-regulated in hepatocellular carcinoma, URHC)，但其是否充当miRNA海绵（Sponge）来调控其目的基因的表达参与HCC的发生发展尚不明确。因此我们研究URHC充当miRNA海绵调控其下游靶基因对HCC恶性表型影响的分子机制。本课题通过荧光原位杂交实验证实URHC定位在细胞质中；生物信息学预测URHC可能调控的miRNA（miR-5007-3p）及miRNA调控的潜在靶基因（DNAJB9），RT-qPCR和荧光素酶报告基因实验分析三者的相关性；Rescue实验进一步证实三者相关性；体内裸鼠成瘤实验分析URHC通过miR-5007-3p及其靶基因对HCC生长具有促进作用。研究表明URHC作为miR-5007-3p海绵（Sponge）调控其目的基因DNAJB9表达参与HCC细胞的生长，从而参与HCC的发生发展。这将有助于阐明URHC充当miRNA海绵（Sponge）介导HCC生长的分子机制，为临床肝癌的诊断、预后及预防提供有效的生物学标志和潜在靶点。