LncRNA-HGBC stabilized by HuR promotes gallbladder cancer progression by regulating miR-502-3p/SET/AKT axis
LncRNA-HGBC stabilized by HuR promotes gallbladder cancer progression by regulating miR-502-3p/SET/AKT axis
复制标题
HuR 稳定的 LncRNA-HGBC 通过调节 miR-502-3p/SET/AKT 轴促进胆囊癌进展
DOI：
10.1186/s12943-019-1097-9
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发表时间：
2019-11-21
期刊：
影响因子：
37.3
通讯作者：
Liu, Ying-bin
中科院分区：
文献类型：
--
作者：
Hu, Yun-ping;Jin, Yun-peng;Liu, Ying-bin
研究背景长链非编码RNA（longnoncodingRNAs，lncRNA）是一类重要的调控因子，通过多种分子机制调控肿瘤的发生和转移。然而，lncRNA在胆囊癌（GBC）中的病理作用仍然很大程度上未知。本研究发现了一种新的lncRNA--lncRNA-HGBC（Highly expressed in GBC），该基因在GBC组织中表达上调，旨在探讨其在GBC发生发展中的作用及其调控机制。通过5′和3′端快速扩增技术（RACE）获得lncRNA-HGBC全长。采用荧光原位杂交（FISH）和亚细胞分级分离法检测lncRNA-HGBC的细胞定位。进行体外和体内测定以探索lncRNA-HGBC在GBC细胞中的生物学效应。RNA pull-down分析、质谱和RNA免疫沉淀（RIP）分析用于鉴定lncRNA-HGBC相互作用蛋白。结果lncRNA-HGBC在GBC中表达上调，其表达上调可预测GBC的生存率。在GBC细胞系中，lncRNA-HGBC的过表达或敲低分别导致体外和异种移植肿瘤中细胞增殖和侵袭的增加或减少。lncRNA-HGBC特异性结合RNA结合蛋白Hu抗原R（HuR），进而稳定lncRNA-HGBC。LncRNA-HGBC作为竞争性内源性RNA与抑制靶基因SET的miR-502- 3 p结合。lncRNA-HGBC的过表达、敲低或突变改变了miR-502- 3 p对SET表达和下游AKT激活的抑制作用。临床上，lncRNA-HGBC表达与miR-502- 3 p呈负相关，与SET和HuR呈正相关。结论lncRNA-HGBC通过激活miR-502- 3 p-SET-AKT级联反应促进GBC转移，提示lncRNA-HGBC是一种新的预后预测因子和治疗靶点。
BackgroundsLong non-coding RNAs (lncRNAs) are essential factors that regulate tumor development and metastasis via diverse molecular mechanisms in a broad type of cancers. However, the pathological roles of lncRNAs in gallbladder carcinoma (GBC) remain largely unknown. Here we discovered a novel lncRNA termed lncRNA Highly expressed in GBC (lncRNA-HGBC) which was upregulated in GBC tissue and aimed to investigate its role and regulatory mechanism in the development and progression of GBC.MethodsThe expression level of lncRNA-HGBC in GBC tissue and different cell lines was determined by quantitative real-time PCR. The full length of lncRNA-HGBC was obtained by 5′ and 3′ rapid amplification of the cDNA ends (RACE). Cellular localization of lncRNA-HGBC was detected by fluorescence in situ hybridization (FISH) assays and subcellular fractionation assay. In vitro and in vivo assays were preformed to explore the biological effects of lncRNA-HGBC in GBC cells. RNA pull-down assay, mass spectrometry, and RNA immunoprecipitation (RIP) assay were used to identify lncRNA-HGBC-interacting proteins. Dual luciferase reporter assays, AGO2-RIP, and MS2-RIP assays were performed to verify the interaction between lncRNA-HGBC and miR-502-3p.ResultsWe found that lncRNA-HGBC was upregulated in GBC and its upregulation could predict poor survival. Overexpression or knockdown of lncRNA-HGBC in GBC cell lines resulted in increased or decreased, respectively, cell proliferation and invasion in vitro and in xenografted tumors. LncRNA-HGBC specifically bound to RNA binding protein Hu Antigen R (HuR) that in turn stabilized lncRNA-HGBC. LncRNA-HGBC functioned as a competitive endogenous RNA to bind to miR-502-3p that inhibits target gene SET. Overexpression, knockdown or mutation of lncRNA-HGBC altered the inhibitory effects of miR-502-3p on SET expression and downstream activation of AKT. Clinically, lncRNA-HGBC expression was negatively correlated with miR-502-3p, but positively correlated with SET and HuR in GBC tissue.ConclusionsOur study demonstrates that lncRNA-HGBC promotes GBC metastasis via activation of the miR-502-3p-SET-AKT cascade, pointing to lncRNA-HGBC as a new prognostic predictor and a therapeutic target.