miR-200bc/429 cluster targets PLCγ1 and differentially regulates proliferation and EGF-driven invasion than miR-200a/141 in breast cancer
miR-200bc/429 cluster targets PLCγ1 and differentially regulates proliferation and EGF-driven invasion than miR-200a/141 in breast cancer
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DOI：
10.1038/onc.2010.201
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发表时间：
2010-07-01
期刊：
影响因子：
8
通讯作者：
Sahin, Oe
中科院分区：
文献类型：
--
作者：
Uhlmann, S.;Zhang, J. D.;Sahin, Oe
编码人类miR-200家族的microRNA的基因映射到脆性染色体区域，并且在肿瘤进展时经常下调。虽然已经报道了调节上皮-间质转化和转化生长因子-β驱动的细胞侵袭，但miR-200家族在EGF驱动的乳腺癌细胞侵袭、存活力、凋亡和细胞周期进展中的作用仍然未知。特别是，没有研究比较该miRNA家族的两个簇的作用。在这项研究中，我们首次发现miR-200家族成员差异调节EGF驱动的乳腺癌细胞的侵袭、存活、凋亡和细胞周期进程。我们发现，所有miR-200家族成员都调节EGF驱动的侵袭，其中miR-200 bc/429簇比miR-200 a/141簇显示出更强的作用。此外，miR-200 a/141簇的表达导致G1期阻滞，这得到了p27/Kip 1表达增加和细胞周期蛋白依赖性激酶6表达减少的支持。相反，200 bc/429簇的表达减少了G1群体并增加了G2/M期，分别与观察到的p27/Kip 1的减少和Cdc 25 C的抑制性磷酸化的上调一致。为了验证这一假设，即两个簇之间观察到的表型差异是由差异靶向光谱引起的，我们结合功能获得研究进行了全基因组微阵列分析。这鉴定了磷脂酶C γ 1（PLCG 1），其仅被miR-200 bc/429簇下调，作为促成这些表型差异的潜在候选者。荧光素酶报告基因测定验证了PLCG 1是miR-200 bc/429簇的直接功能靶点，但不是miR-200 a/141簇的直接功能靶点。最后，PLCG 1的缺失部分模拟了miR-200 bc/429过表达在乳腺癌细胞的存活率、凋亡和EGF驱动的细胞侵袭中的作用。我们的研究结果表明，miR-200家族具有肿瘤抑制功能，通过负调节EGF驱动的细胞侵袭，生存力和细胞周期进程在乳腺癌中。Oncogene（2010）29，4297-4306; doi：10.1038/onc.2010.201; 2010年5月31日在线发表
The genes encoding microRNAs of the human miR-200 family map to fragile chromosomal regions and are frequently downregulated upon tumor progression. Although having been reported to regulate epithelial-to-mesenchymal transition and transforming growth factor-beta- driven cell invasion, the role of the miR-200 family in EGF-driven breast cancer cell invasion, viability, apoptosis and cell cycle progression is still unknown. In particular, there is no study comparing the roles of the two clusters of this miRNA family. In this study, we show for the first time that miR-200 family members differentially regulate EGF-driven invasion, viability, apoptosis and cell cycle progression of breast cancer cells. We showed that, all miR-200 family members regulate EGF-driven invasion, with the miR-200bc/429 cluster showing stronger effects than the miR-200a/141 cluster. Furthermore, expression of the miR-200a/141 cluster results in G1 arrest supported by increased p27/Kip1 and decreased cyclin dependent kinase 6 expression. In contrast, expression of the 200bc/429 cluster decreases G1 population and increases G2/M phase, in line with the observed reduction of p27/Kip1 and upregulation of the inhibitory phosphorylation of Cdc25C, respectively. To test the hypothesis that phenotypical differences observed between the two clusters are caused by differential targeting spectrums, we performed genome-wide microarray profiling in combination with gain-of-function studies. This identified phospholipase C gamma 1 (PLCG1), which was downregulated only by the miR-200bc/429 cluster, as a potential candidate contributing to these phenotypical differences. Luciferase reporter assays validated PLCG1 as a direct functional target of miR-200bc/429 cluster, but not of miR-200a/141 cluster. Finally, loss of PLCG1 in part mimicked the effect of miR-200bc/429 overexpression in viability, apoptosis and EGF-driven cell invasion of breast cancer cells. Our results suggest that the miR-200 family has a tumor-suppressor function by negatively regulating EGF-driven cell invasion, viability and cell cycle progression in breast cancer. Oncogene (2010) 29, 4297-4306; doi: 10.1038/onc.2010.201; published online 31 May 2010