Ankyrin repeat and single KH domain 1 (ANKHD1) drives renal cancer cell proliferation via binding to and altering a subset of miRNAs
Ankyrin repeat and single KH domain 1 (ANKHD1) drives renal cancer cell proliferation via binding to and altering a subset of miRNAs
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DOI：
10.1074/jbc.ra117.000975
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发表时间：
2018-06-22
影响因子：
4.8
通讯作者：
Zeidler, Martin P.
中科院分区：
文献类型：
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作者：
Fragiadaki, Maria;Zeidler, Martin P.
透明细胞肾细胞癌（ccRCC）是全球最常见的肾癌。与异常microRNA（miRNA）调节相关的细胞增殖增加是致癌的标志。锚蛋白重复序列和单KH结构域1（ANKHD 1）是一种高度保守的蛋白质，被发现与果蝇中的核心癌症通路相互作用;然而，它在RCC中的参与完全未被探索。结合大规模基因组学数据集的定量PCR研究表明，与健康对照相比，ANKHD 1在RCC患者的肾脏中显著上调。细胞周期分析显示ANKHD 1是肾细胞癌细胞分裂的关键因子。为了了解ANKHD 1驱动增殖的分子机制，我们进行了生物信息学分析，结果显示ANKHD 1含有一个假定的miRNA结合基序。我们筛选了48种具有肿瘤增强或抑制活性的miRNA，发现ANKHD 1结合并调节三种肿瘤抑制miRNA（即miR-29 a，miR-205和miR-196 a）。RNA免疫沉淀分析表明，ANKHD 1通过位于蛋白质C末端的单个K-同源结构域与其靶miRNA发生物理相互作用。在功能上，我们发现ANKHD 1通过结合并主要抑制miR-29 a以及通过miR-196 a/205在较小程度上积极驱动ccRCC细胞有丝分裂，导致增殖基因如CCDN 1的上调。总的来说，这些数据将ANKHD 1鉴定为通过特异性miRNA相互作用的ccRCC增殖的新调节因子。
Clear cell renal cell carcinoma (ccRCC) represents the most common kidney cancer worldwide. Increased cell proliferation associated with abnormal microRNA (miRNA) regulation are hallmarks of carcinogenesis. Ankyrin repeat and single KH domain 1 (ANKHD1) is a highly conserved protein found to interact with core cancer pathways in Drosophila; however, its involvement in RCC is completely unexplored. Quantitative PCR studies coupled with large-scale genomics data sets demonstrated that ANKHD1 is significantly up-regulated in kidneys of RCC patients when compared with healthy controls. Cell cycle analysis revealed that ANKHD1 is an essential factor for RCC cell division. To understand the molecular mechanism(s) utilized by ANKHD1 to drive proliferation, we performed bioinformatics analyses that revealed that ANKHD1 contains a putative miRNA-binding motif. We screened 48 miRNAs with tumor-enhancing or -suppressing activities and found that ANKHD1 binds to and regulates three tumor-suppressing miRNAs (i.e. miR-29a, miR-205, and miR-196a). RNA-immunoprecipitation assays demonstrated that ANKHD1 physically interacts with its target miRNAs via a single K-homology domain, located in the C terminus of the protein. Functionally, we discovered that ANKHD1 positively drives ccRCC cell mitosis via binding to and suppressing mainly miR-29a and to a lesser degree via miR-196a/205, leading to up-regulation in proliferative genes such as CCDN1. Collectively, these data identify ANKHD1 as a new regulator of ccRCC proliferation via specific miRNA interactions.