From the Academic Nephrology Unit, Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield S10 2RX, United Kingdom and
the Bateson Centre, Departments of Biomedical Science, University of Sheffield, Sheffield S10 2TN, United Kingdom.
J Biol Chem. 2018 Jun 22;293(25):9570-9579. doi: 10.1074/jbc.RA117.000975. Epub 2018 Apr 25.
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 ; 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 ( 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.
透明细胞肾细胞癌 (ccRCC) 是全球最常见的肾癌。与异常 microRNA (miRNA) 调控相关的细胞增殖增加是癌变的标志。锚蛋白重复和单个 KH 结构域 1 (ANKHD1) 是一种高度保守的蛋白质，已被发现与核心癌症途径相互作用；然而，它在 RCC 中的参与是完全未知的。定量 PCR 研究结合大规模基因组数据集表明，与健康对照相比，ANKHD1 在 RCC 患者的肾脏中显著上调。细胞周期分析显示，ANKHD1 是 RCC 细胞分裂的必需因素。为了了解 ANKHD1 用于驱动增殖的分子机制，我们进行了生物信息学分析，结果表明 ANKHD1 含有一个假定的 miRNA 结合基序。我们筛选了具有肿瘤增强或抑制活性的 48 种 miRNA，发现 ANKHD1 与三个肿瘤抑制 miRNA (miR-29a、miR-205 和 miR-196a) 结合并调节它们。RNA 免疫沉淀试验表明，ANKHD1 通过位于蛋白质 C 末端的单个 KH 结构域与靶 miRNA 发生物理相互作用。功能上，我们发现 ANKHD1 通过与 miR-29a 结合并抑制其主要作用，以及通过 miR-196a/205 产生次要作用，从而正向驱动 ccRCC 细胞有丝分裂，导致细胞周期蛋白 D1 (CCDN1) 等增殖基因上调。总的来说，这些数据表明 ANKHD1 是通过特定的 miRNA 相互作用调节 ccRCC 增殖的新调节因子。
