The tumor-suppressive miR-497-195 cluster targets multiple cell-cycle regulators in hepatocellular carcinoma.
The tumor-suppressive miR-497-195 cluster targets multiple cell-cycle regulators in hepatocellular carcinoma.
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DOI：
10.1371/journal.pone.0060155
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Inazawa J
中科院分区：
文献类型：
--
作者：
Furuta M;Kozaki K;Tanimoto K;Tanaka S;Arii S;Shimamura T;Niida A;Miyano S;Inazawa J
MicroRNAs(MiRNAs)是基因表达的关键转录后调节因子，在肿瘤发生过程中常被解除调控。为了探索在肝细胞癌中具有重要功能的肿瘤抑制(TS)-miRNAs，我们在六个肝癌细胞系中进行了基于功能和表达的TS-miRNAs的综合筛选。筛选发现了7个miRNA，它们通过每个miRNA的过表达而显示出抑制生长的活性，并且在肝癌细胞系中内源性下调。对一大批肝癌细胞系和原发肿瘤的进一步表达分析表明，四个miRNAs，miR-101，-195，-378和-497，作为候选的TS-miRNAs经常在肝癌中沉默。其中，miR-195和miR-497两个聚集型miRNAs具有明显的生长抑制活性，并诱导细胞周期停滞。利用Argonute2-免疫沉淀-深度测序(Ago2-IP-seq)和全基因组表达谱对它们的靶标进行了全面的探索，在它们过表达之后进行了途径分析，显示出细胞周期调节因子的显著丰富。在候选基因中，我们成功地确定了CCNE1、CDC25A、CCND3、CDK4和BTRC作为miR-497和miR-195的直接靶点。此外，肿瘤特异性上调的靶基因CDK6、CCNE1、CDC25A和CDK4与miR-195和miR-497的表达呈负相关。这些结果表明，通过下调miR-195和miR-497的表达，调控细胞周期进程的分子途径发生了整体改变，导致了肝癌发生过程中细胞的异常增殖。
MicroRNAs (miRNAs) are key post-transcriptional regulators of gene expression and commonly deregulated in carcinogenesis. To explore functionally crucial tumor-suppressive (TS)-miRNAs in hepatocellular carcinoma (HCC), we performed integrative function- and expression-based screenings of TS-miRNAs in six HCC cell lines. The screenings identified seven miRNAs, which showed growth-suppressive activities through the overexpression of each miRNA and were endogenously downregulated in HCC cell lines. Further expression analyses using a large panel of HCC cell lines and primary tumors demonstrated four miRNAs, miR-101, -195, -378 and -497, as candidate TS-miRNAs frequently silenced in HCCs. Among them, two clustered miRNAs miR-195 and miR-497 showed significant growth-suppressive activity with induction of G1 arrest. Comprehensive exploration of their targets using Argonute2-immunoprecipitation-deep-sequencing (Ago2-IP-seq) and genome-wide expression profiling after their overexpression followed by pathway analysis, revealed a significant enrichment of cell cycle regulators. Among the candidates, we successfully identified CCNE1, CDC25A, CCND3, CDK4, and BTRC as direct targets for miR-497 and miR-195. Moreover, target genes frequently upregulated in HCC in a tumor-specific manner, such as CDK6, CCNE1, CDC25A and CDK4, showed an inverse correlation in the expression of miR-195 and miR-497, and their targets. These results suggest the molecular pathway regulating cell cycle progression to be integrally altered by downregulation of miR-195 and miR-497 expression, leading to the aberrant cell proliferation in hepatocarcinogenesis.
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影响因子：
5.6
作者：
Finnerty JR;Wang WX;Hébert SS;Wilfred BR;Mao G;Nelson PT
通讯作者：
Nelson PT
影响因子：
13.6
作者：
Di Leva G;Croce CM
通讯作者：
Croce CM
DOI：
10.1111/j.1440-1746.2008.05666.x
发表时间：
2009-04-01
影响因子：
4.1
作者：
Guo, Junming;Miao, Ying;Wang, Yanjun
通讯作者：
Wang, Yanjun
影响因子：
13.5
作者：
Song, Guisheng;Sharma, Amar Deep;Roll, Garrett R.;Ng, Raymond;Lee, Andrew Y.;Blelloch, Robert H.;Frandsen, Niels M.;Willenbring, Holger
通讯作者：
Willenbring, Holger
DOI：
10.1073/pnas.0706901104
发表时间：
2007-12-18
影响因子：
11.1
作者：
Lee, Daniel Y.;Deng, Zhaoqun;Yang, Burton B.
通讯作者：
Yang, Burton B.