microRNA-26a and-584 inhibit the colorectal cancer progression through inhibition of the binding of hnRNP A1-CDK6 mRNA
microRNA-26a and-584 inhibit the colorectal cancer progression through inhibition of the binding of hnRNP A1-CDK6 mRNA
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DOI：
10.1016/j.bbrc.2015.10.055
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发表时间：
2015-11-27
影响因子：
3.1
通讯作者：
Kohgo, Yutaka
中科院分区：
文献类型：
--
作者：
Konishi, Hiroaki;Fujiya, Mikihiro;Kohgo, Yutaka
虽然化疗和分子靶向治疗的进展改善了结直肠癌患者的预后，但结肠癌的死亡率仍然很高，这表明需要开发新的治疗靶点来改善结肠癌的预后。异质性核糖核蛋白A1（hnRNPA 1）在大肠癌中高表达，其表达与大肠癌的恶性转化有关。在这项研究中，我们用RNA免疫沉淀（RNA-IP）方法进行了微阵列分析，并在结肠直肠癌细胞系SW 620中鉴定了hnRNP A1相互作用的miR，包括miR-26 a和-584。SRB测定揭示了miR-26 a和-584的肿瘤抑制作用，并且这些miR的肿瘤抑制作用通过hnRNPA 1的下调而减弱。转录组分析和RNA-IP的组合方法揭示了hnRNP A1相互作用的mRNA，包括细胞周期蛋白依赖性激酶6（CDK 6）。Western印迹分析揭示了miR-26 a和-584过表达细胞以及hnRNP A1敲低细胞中CDK 6的下调。结合实验表明，转染miR-26 a和-584后，hnRNP A1-CDK 6 mRNA的结合能力降低。在miR-26 a和-584过表达细胞中诱导切割的caspase-3的表达。这些数据表明miR-26 a和-584抑制hnRNP A1-CDK 6 mRNA的结合并诱导结直肠癌细胞凋亡。(C)2015 Elsevier Inc. All rights reserved.
While the progress of chemotherapy and molecular targeted therapy has improved the outcome of colorectal cancer patients, the mortality of colon cancer remains high, indicating the need to develop novel therapeutic targets for improving the outcome of colon cancer. Heterogeneous ribonucleoprotein A1 (hnRNP A1) is highly expressed in colorectal cancer and its expression correlates with malignant transformation. In this study, we performed a microarray analysis with the RNA immunoprecipitation (RNA-IP) method and identified hnRNP A1-interacting miRs, including miR-26a and -584, in a colorectal cancer cell line, SW620. A SRB assay revealed the tumor suppressive effect of miR-26a and -584, and the tumor suppressive effect of these miRs was diminished by the downregulation of hnRNP A1. The combined method of a transcriptome analysis and RNA-IP revealed hnRNP A1-interacting mRNAs, including cyclin dependent kinase 6 (CDK6). A Western blot analysis revealed the downregulation of CDK6 in miR-26a and -584 overexpression cells, as well as hnRNP A1 knockdown cells. The binding assay indicated that the binding of hnRNP A1-CDK6 mRNA was reduced by transfection of miR-26a and -584. The expression of cleaved caspase-3 was induced in miR-26a and -584 overexpression cells. These data indicate that miR-26a and -584 inhibit the binding of hnRNP A1-CDK6 mRNA and induce colorectal cancer cell apoptosis. (C) 2015 Elsevier Inc. All rights reserved.