miR-214/199a/199a* cluster levels predict poor survival in hepatocellular carcinoma through interference with cell-cycle regulators.
miR-214/199a/199a* cluster levels predict poor survival in hepatocellular carcinoma through interference with cell-cycle regulators.
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miR-214/199a/199acluster 水平通过干扰细胞周期调节因子预测肝细胞癌的较差生存率
DOI：
10.18632/oncotarget.6137
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发表时间：
2016-01-05
期刊：
影响因子：
--
通讯作者：
Xie C
中科院分区：
文献类型：
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作者：
Wang P;Chen S;Fang H;Wu X;Chen D;Peng L;Gao Z;Xie C
探讨miR-214/199 a/199 a * 簇在人肝细胞癌（HCC）中的临床和功能相关性，阐明miR-214的作用机制。Kaplan-Meier和考克斯比例回归分析用于确定miR-214/199 a/199 a * 簇水平与HCC患者生存率的相关性。用miR-214模拟物/受体处理的细胞研究miR-214在调节HCC细胞增殖中的作用。此外，在具有miR-214模拟物/抑制剂以增加/减少miR-214表达的HCC细胞系中评估miR-214对E2 F2、细胞周期蛋白依赖性激酶（CDK）3和CDK 6表达的抑制作用。通过双荧光素酶报告基因分析验证miR-214与E2 F2、CDK 3和CDK 6的3′-非翻译区的直接结合。在分析HCC临床标本和细胞系时，我们发现与非癌组织或正常肝上皮细胞系相比，miR-214/199 a/199 a * 表达均匀降低。较高的miR-214水平与患者生存率的改善相关。肝癌细胞中miR-214的过表达通过诱导G1-S检查点阻滞来抑制增殖。相反，RNA干扰介导的miR-214沉默促进了细胞周期进程，加速了HCC细胞的增殖。E2 F2、CDK 3和CDK 6分别被miR-214直接靶向抑制，恢复它们的表达逆转了miR-214对细胞周期进程的抑制。miR-214的表达与其靶点之间的关系在HCC肿瘤异种移植物和临床标本中得到证实。我们的研究结果表明，miR-214通过抑制E2 F2、CDK 3和CDK 6在HCC中具有肿瘤抑制活性。
To identify the clinical and functional association of miR-214/199a/199a* cluster in human hepatocellular carcinoma (HCC) and to clarify the mechanism of miR-214. Kaplan-Meier and Cox proportional regression analyses were used to determine the association of miR-214/199a/199a* cluster levels with the survival of HCC patients. The role of miR-214 in regulating HCC cell proliferation was studied with miR-214 mimics/inhibitor-treated cells. Furthermore, the inhibition effect of miR-214 on E2F2, cyclin-dependent kinase (CDK) 3 and CDK6 expression was assessed in HCC cell lines with miR-214 mimics/inhibitors to increase/decrease miR-214 expression. Direct binding of miR-214 to the 3′-untranslated regions of E2F2, CDK3, and CDK6 was verified by dual-luciferase reporter assay. In analyzing HCC clinical specimens and cell lines, we discovered a uniform decrease in miR-214/199a/199a* expression in comparison with noncancerous tissue or normal liver epithelial cell lines. Higher miR-214 levels were related with improved patient survival. Overexpression of miR-214 in HCC cells inhibited proliferation by inducing G1-S checkpoint arrest. Conversely, RNA interference–mediated silencing of miR-214 promoted cell-cycle progression and accelerated the proliferation of HCC cells. E2F2, CDK3 and CDK6 were each directly targeted for inhibition by miR-214, and restoring their expression reversed miR-214 inhibition of cell-cycle progression. The relationship between expression of miR-214 and its targets was confirmed in HCC tumor xenografts and clinical specimens. Our results demonstrate that miR-214 has tumor-suppressive activity in HCC through inhibition of E2F2, CDK3 and CDK6.