HOTAIR Long Noncoding RNA Promotes Gastric Cancer Metastasis through Suppression of Poly r(C)-Binding Protein (PCBP) 1
HOTAIR Long Noncoding RNA Promotes Gastric Cancer Metastasis through Suppression of Poly r(C)-Binding Protein (PCBP) 1
复制标题
HOTAIR 长非编码 RNA 通过抑制聚 r(c) 结合蛋白 (PCBP) 1 促进胃癌转移
DOI：
10.1158/1535-7163.mct-14-0695
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发表时间：
2015-05-01
影响因子：
5.7
通讯作者：
Xu, Jia
中科院分区：
文献类型：
--
作者：
Zhang, Zi-Zhen;Shen, Zhi-Yong;Xu, Jia
本研究的目的是评估HOTAIR长链非编码RNA在胃癌转移中的作用。我们通过实时逆转录PCR和北方印迹分析100例胃组织（50例胃癌组织和50例癌旁正常粘膜）和4种胃癌细胞系中HOTAIR的表达水平。进行瞬时RNAi介导的HOTAIR敲低和pcDNA介导的HOTAIR过表达。在异种移植试验中，通过稳定的shRNA介导的HOTAIR敲减和慢病毒介导的HOTAIR过表达来研究HOTAIR对体内致瘤性和转移负荷的作用。进行蛋白质组学分析以解读具有不同HOTAIR表达水平的细胞中的差异蛋白质表达。其中一种差异调节蛋白Poly r（C）结合蛋白（PCBP）1随后得到验证，并通过异种移植试验评估其功能。HOTAIR在癌组织中的表达显著高于癌旁正常粘膜。HOTAIR表达水平决定了这些细胞的体外和体内致瘤性和转移潜力。PCBP1和HOTAIR在表达水平和功能上都具有相反的关系。通过RNA免疫沉淀结合定量真实的时间PCR证实了两者之间的直接相互作用。PCBP1被证实是胃癌发病机制的抑制剂，并且在功能上与HOTAIR长非编码RNA相反。因此，HOTAIR表达可能作为一个潜在的重要的疾病生物标志物，用于识别高危胃癌患者。此外，我们的研究结果为HOTAIR过表达和PCBP1下调以及培养和异种移植胃癌细胞中随后的恶性表型提供了机制证据。(C)2015年AACR。
The objective of this study was to evaluate the role of HOTAIR long noncoding RNA in gastric cancer metastasis. We analyzed HOTAIR expression levels by real-time reverse transcription PCR and Northern blot analysis in 100 gastric tissues (50 gastric cancer tissues and 50 adjacent normal mucosa), and in four gastric cancer cell lines. Transient RNAi-mediated knockdown and pcDNA-mediated overexpression of HOTAIR were performed. Stable shRNA-mediated knockdown and lentiviral-mediated overexpression of HOTAIR were to study the role of HOTAIR on in vivo tumorigenicity and metastatic burden in the context of xenograft assays. Proteomic profiling was performed to decipher differential protein expression in cells with different HOTAIR expression levels. One of the differentially regulated proteins, Poly r(C)binding protein (PCBP) 1, was subsequently validated and its function evaluated through xenograft assays. Expression of HOTAIR was significantly higher in cancerous tissues than in adjacent normal mucosa. HOTAIR expression levels dictated in vitro and in vivo tumorigenicity and metastatic potential in these cells. PCBP1 and HOTAIR have an inverse relationship, both at expression level and in function. A direct interaction between the two was confirmed through RNA immunoprecipitation coupled with quantitative real- time PCR. PCBP1 was confirmed to be an inhibitor of gastric cancer pathogenesis and as functionally opposite to HOTAIR long noncoding RNA. In conclusion, HOTAIR expression may serve as a potentially important disease biomarker for the identification of high-risk gastric cancer patients. Moreover, our findings provide mechanistic evidence for HOTAIR overexpression and PCBP1 downregulation and the ensuing malignant phenotype in both cultured and xenograft gastric cancer cells. (C) 2015 AACR.