Long Noncoding RNA GCASPC, a Target of miR-17-3p, Negatively Regulates Pyruvate Carboxylase-Dependent Cell Proliferation in Gallbladder Cancer
Long Noncoding RNA GCASPC, a Target of miR-17-3p, Negatively Regulates Pyruvate Carboxylase-Dependent Cell Proliferation in Gallbladder Cancer
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长非编码 RNA GCASPC（miR-17-3p 的靶标）负调节胆囊癌中丙酮酸羧化酶依赖性细胞增殖
DOI：
10.1158/0008-5472.can-15-3047
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发表时间：
2016-09-15
期刊：
影响因子：
11.2
通讯作者：
Quan, Zhi-wei
中科院分区：
文献类型：
--
作者：
Ma, Ming-zhe;Zhang, Yan;Quan, Zhi-wei
长链非编码rna （lncRNA）与许多癌症的发展有关。在这里，我们报告了lncRNA GCASPC在决定胆囊癌进展中的关键作用的发现。通过表达微阵列分析，鉴定并验证了5例胆囊癌标本和配对邻近非肿瘤组织之间差异表达的lncrna和mrna。采用实时荧光定量PCR检测42例胆囊癌患者组织中GCASPC水平，并在89例胆囊癌患者的单独队列中进一步证实GCASPC水平。GCASPC在几种胆囊癌细胞系中过表达或沉默，并进行了分子和生物学分析。胆囊癌中的GCASPC水平明显低于邻近的非肿瘤组织，并与肿瘤大小、美国癌症联合委员会肿瘤分期和患者预后相关。GCASPC过表达可抑制体外和体内细胞增殖，而GCASPC沉默则具有相反的作用。通过RNA pull-down和质谱分析，我们确定丙酮酸羧化酶是与GCASPC相关的RNA结合蛋白。由于GCASPC是miR-17-3p的靶标，我们证实miR-17-3p和GCASPC都通过限制蛋白质稳定性下调丙酮酸羧化酶的水平和活性。综上所述，我们的研究结果确定了lncrna调控的胆囊癌细胞增殖的新机制，为理解其致病性提供了新的基础。(c) 2016年aacr。
Long noncoding RNAs (lncRNA) are being implicated in the development of many cancers. Here, we report the discovery of a critical role for the lncRNA GCASPC in determining the progression of gallbladder cancer. Differentially expressed lncRNAs and mRNAs between gallbladder cancer specimens and paired adjacent nontumor tissues from five patients were identified and validated by an expression microarray analysis. Quantitative real-time PCR was used to measure GCASPC levels in tissues from 42 gallbladder cancer patients, and levels of GCASPC were confirmed further in a separate cohort of 89 gallbladder cancer patients. GCASPC was overexpressed or silenced in several gallbladder cancer cell lines where molecular and biological analyses were performed. GCASPC levels were significantly lower in gallbladder cancer than adjacent nontumor tissues and were associated with tumor size, American Joint Committee on Cancer tumor stage, and patient outcomes. GCASPC overexpression suppressed cell proliferation in vitro and in vivo, whereas GCASPC silencing had opposite effects. By RNA pull-down and mass spectrometry, we identified pyruvate carboxylase as an RNAbinding protein that associated with GCASPC. Because GCASPC is a target of miR-17-3p, we confirmed that both miR-17-3p and GCASPC downregulated pyruvate carboxylase level and activity by limiting protein stability. Taken together, our results defined a novel mechanism of lncRNA-regulated cell proliferation in gallbladder cancer, illuminating a new basis for understanding its pathogenicity. (C) 2016 AACR.