Overexpression of miR-210, a downstream target of HIF1α, causes centrosome amplification in renal carcinoma cells
Overexpression of miR-210, a downstream target of HIF1α, causes centrosome amplification in renal carcinoma cells
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DOI：
10.1002/path.2860
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发表时间：
2011-06-01
影响因子：
7.3
通讯作者：
Moriyama, Masatsugu
中科院分区：
文献类型：
--
作者：
Nakada, Chisato;Tsukamoto, Yoshiyuki;Moriyama, Masatsugu
MiR-210在透明细胞肾细胞癌（CCC）中显著上调，但MiR-210上调的机制和生物学后果尚不清楚。在这里，我们发现miR-210在肾癌细胞系中高表达，其表达与缺氧和缺氧条件下缺氧诱导因子1 α (HIF1 α)的积累明显相关，提示肾癌细胞中miR-210的上调很可能是由于HIF1 α的积累。为了揭示miR-210上调的作用，我们将miR-210前体转染到肾癌细胞中。转染后，细胞聚集在细胞周期的G2/M期，细胞活力下降，提示miR-210过表达可能引发阻碍正常细胞分裂的事件。免疫细胞化学表明，在过表达miR-210的细胞中，多极纺锤体伴随着中心体扩增。有报道称，中心体扩增引起染色体错分离，最终导致染色体不稳定和非整倍体。事实上，在miR-210过表达的细胞中，非整倍体细胞（bbb4n）的比例增加。通过TargetScan和PicTar算法，E2F3被确定为miR-210的可能靶点之一，并在蛋白水平上被miR-210抑制。此外，转染E2F3 siRNA的细胞中非整倍体细胞的比例增加。基于这些结果，我们提出，HIF1 α积累导致的miR-210上调可能至少部分通过下调E2F3诱导非整倍体，并可能在CCC的肿瘤发生和/或进展中发挥作用。版权所有(C) 2011英国和爱尔兰病理学会。约翰·威利父子有限公司出版。
MiR-210 is significantly up-regulated in clear cell renal cell carcinoma (CCC), but the mechanism and biological consequences of miR-210 up-regulation are poorly understood. Here, we show that miR-210 is highly expressed in renal carcinoma cell lines and that its expression is clearly correlated with accumulation of hypoxia-inducible factor 1 alpha (HIF1 alpha) under normoxia as well as hypoxia, suggesting that miR-210 up-regulation in renal carcinoma cells is most likely due to accumulation of HIF1 alpha. To reveal the effects of miR-210 up-regulation, the miR-210 precursor was transfected into renal carcinoma cells. After transfection, the cells accumulated at the G2/M phase of the cell cycle and their viability was decreased, suggesting that miR-210 overexpression may trigger an event that hinders normal cell division. Immunocytochemistry demonstrated a multipolar spindle accompanied by centrosome amplification in cells overexpressing miR-210. It has been reported that centrosome amplification induces chromosome mis-segregation, finally leading to chromosome instability and aneuploidy. Indeed, the proportion of aneuploid cells (>4n) was increased in miR-210 overexpressed cells. By using the TargetScan and PicTar algorithms, E2F3 was identified as one of the possible targets of miR-210 and was suppressed at the protein level by miR-210. Moreover, the proportion of aneuploid cells was increased in E2F3 siRNA transfected cells. On the basis of these results, we propose that miR-210 up-regulation due to HIF1 alpha accumulation may induce aneuploidy via E2F3 down-regulation at least in part, and may play a role in tumourigenesis and/or progression of CCC. Copyright (C) 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.