Long Non-Coding RNA LUCAT1 Promotes Proliferation and Invasion in Clear Cell Renal Cell Carcinoma Through AKT/GSK-3β Signaling Pathway
Long Non-Coding RNA LUCAT1 Promotes Proliferation and Invasion in Clear Cell Renal Cell Carcinoma Through AKT/GSK-3β Signaling Pathway
复制标题
长非编码 RNA LUCAT1 通过 AKT/GSK-3β 信号通路促进透明细胞肾细胞癌的增殖和侵袭
DOI：
10.1159/000491957
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Xie, Wenlian
中科院分区：
文献类型：
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作者：
Zheng, Zaosong;Zhao, Fengjin;Xie, Wenlian
背景/目的：长链非编码rna （IncRNAs）已成为多种癌症的新调控因子和生物标志物。然而，很少有incrna在透明细胞肾细胞癌（ccRCC）中得到很好的表征。方法：采用微阵列分析方法研究5例ccRCC组织及邻近正常组织的IncRNA表达谱。采用实时荧光定量PCR检测90对ccRCC组织中肺癌相关转录本1 （LUCAT1）的表达，并通过The cancer Genome Atlas （TCGA）数据库进行验证。采用Kaplan-Meier分析检验LUCAT1和CXCL2在ccRCC患者中的预后价值。通过功能的丧失和获得来探讨LUCAT1对ccRCC细胞增殖和侵袭的影响。Western blotting评估LUCAT1在ccRCC进展中的潜在机制。通过趋化因子刺激实验研究控制LUCAT1在ccRCC细胞中表达的可能机制。采用酶联免疫吸附法测定ccRCC患者和健康志愿者的血清CXCL2。采用受试者工作特征曲线分析，探讨血清CXCL2对ccRCC的临床诊断价值。结果：我们发现，与正常组织相比，LUCAT1在临床ccRCC组织（n = 90）和TCGA ccRCC组织（n = 448）中均显著上调。统计分析显示，LUCAT1表达水平与肿瘤T分期（P < 0.01）、M分期（P < 0.01）、TNM分期（P < 0.01）呈正相关。lucat1高表达组总生存期和无病生存期明显短于低表达组（log-rank P < 0.01）。LUCAT1敲低抑制ccRCC细胞增殖和集落形成，诱导细胞周期阻滞于G1期，抑制细胞迁移和侵袭。LUCAT1的过表达郑铮和赵f对这项工作贡献相同。促进ccRCC细胞的增殖、迁移和侵袭。机制研究表明，LUCAT1通过调节细胞周期蛋白D1、细胞周期蛋白依赖性激酶4和磷酸化的视网膜母细胞瘤转录辅助抑制因子1的表达，诱导细胞周期G1阻滞。此外，LUCAT1在一定程度上通过诱导AKT磷酸化和抑制GSK-3 β磷酸化来促进ccRCC细胞的增殖和侵袭。我们还发现，趋化因子CXCL2在ccRCC中上调，诱导LUCAT1表达，可能是ccRCC的诊断和预后生物标志物。结论：LUCAT1在ccRCC组织及肾癌细胞系中表达上调，且与ccRCC的恶性分期及不良预后显著相关。LUCAT1通过AKT/GSK-3 β信号通路促进ccRCC细胞增殖和侵袭。我们还发现，趋化因子CXCL2可诱导LUCAT1过表达。这些发现表明CXCL2/LUCAT1/AKT/GSK3 β轴是ccRCC潜在的治疗靶点和分子生物标志物。(C) 2018作者：s . Karger AG，巴塞尔出版
Background/Aims: Long non-coding RNAs (IncRNAs) have emerged as new regulators and biomarkers in several cancers. However, few IncRNAs have been well characterized in clear cell renal cell carcinoma (ccRCC). Methods: We investigated the IncRNA expression profile by microarray analysis in 5 corresponding ccRCC tissues and adjacent normal tissues. Lung cancer associated transcript 1 (LUCAT1) expression was examined in 90 paired ccRCC tissues by real-time PCR and validated by The Cancer Genome Atlas (TCGA) database. Kaplan-Meier analysis was used to examine the prognostic value of LUCAT1 and CXCL2 in ccRCC patients. Loss and gain of function were performed to explore the effect of LUCAT1 on proliferation and invasion in ccRCC cells. Western blotting was performed to evaluate the underlying mechanisms of LUCAT1 in ccRCC progression. Chemokine stimulation assay was performed to investigate possible mechanisms controlling LUCAT1 expression in ccRCC cells. Enzyme-linked immunosorbent assays were performed to determine serum CXCL2 in ccRCC patients and healthy volunteers. Receiver operating characteristic curve analysis was performed to examine the clinical diagnostic value of serum CXCL2 in ccRCC. Results: We found that LUCAT1 was significantly upregulated in both clinical ccRCC tissues (n = 90) and TCGA ccRCC tissues (n = 448) compared with normal tissues. Statistical analysis revealed that the LUCAT1 expression level positively correlated with tumor T stage (P < 0.01), M stage (P < 0.01), and TNM stage (P < 0.01). Overall survival and disease-free survival time were significantly shorter in the high-LUCAT1-expression group than in the low-LUCAT1-expression group (log-rank P < 0.01). LUCAT1 knockdown inhibited ccRCC cell proliferation and colony formation, induced cell cycle arrest at G1 phase, and inhibited cell migration and invasion. Overexpression of LUCAT1 Z. Zheng and F. Zhao contributed equally to this work. promoted proliferation, migration, and invasion of ccRCC cells. Mechanistic investigations showed that LUCAT1 induced cell cycle G1 arrest by regulating the expression of cyclin D1, cyclin-dependent kinase 4, and phosphorylated retinoblastoma transcriptional corepressor 1. Moreover, LUCAT1 promoted proliferation and invasion in ccRCC cells partly through inducing the phosphorylation of AKT and suppressing the phosphorylation of GSK-3 beta. We also revealed that chemokine CXCL2, upregulated in ccRCC, induced LUCAT1 expression and might be a diagnostic and prognostic biomarker in ccRCC. Conclusions: LUCAT1 was upregulated in ccRCC tissues and renal cancer cell lines, and significantly correlated with malignant stage and poor prognosis in ccRCC. LUCAT1 promoted proliferation and invasion in ccRCC cells through the AKT/GSK-3 beta signaling pathway. We also revealed that LUCAT1 overexpression was induced by chemokine CXCL2. These findings indicate that the CXCL2/LUCAT1/AKT/GSK3 beta axis is a potential therapeutic target and molecular biomarker for ccRCC. (C) 2018 The Author(s) Published by S. Karger AG, Basel