Dual-strand tumor suppressor miR-193b-3p and-5p inhibit malignant phenotypes of lung cancer by suppressing their common targets
Dual-strand tumor suppressor miR-193b-3p and-5p inhibit malignant phenotypes of lung cancer by suppressing their common targets
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DOI：
10.1042/bsr20190634
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发表时间：
2019-07-12
影响因子：
4
通讯作者：
Kim, Hyeon Ho
中科院分区：
文献类型：
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作者：
Choi, Kyung Hee;Shin, Chang Hoon;Kim, Hyeon Ho
新出现的研究表明，microRNAs(MiRNAs)在癌症的恶性过程中发挥着多种作用，包括增殖和获得转移潜能。使用先前建立的脑转移肺癌模型，通过miRNA微阵列寻找与肺癌恶性相关的差异表达的miRNAs。脑转移肺癌细胞中有25个miRNAs表达下调。其中miR-193B-3P和-5P被选作进一步研究。通过Transwell侵袭、伤口愈合和集落形成实验检测它们在转移潜能和增殖中的作用。采用逆转录定量聚合酶链式反应(RT-qPCR)、Western印迹、ArgAerte 2-RNA免疫沉淀(Ago2-RIP)和报告基因分析等方法，探讨miR-193B-3P和-5P的作用机制。MiR-193B的两条链在脑转移的肺癌细胞和肺癌患者的组织中下调。MiR-193B-3p和-5p的过表达抑制了侵袭和迁移活性，并降低了克隆形成能力。相反，抑制miR-193B-3P或-5P可增加细胞的转移潜能和集落形成能力。Cyclin D1(CCND1)、Ajuba Lim蛋白(AJUBA)和带有EGF样结构域的心脏发育蛋白1(HEG1)是miR-193B-3p和-5p的共同靶基因。报告实验和Ago2-RIP实验表明，这两种miRNAs都直接与靶mRNA的3‘非翻译区(3’UTR)结合。目的基因的敲除降低了原发和转移性肺癌细胞的增殖和转移潜能。我们的结果表明miR-193B是一种双链肿瘤抑制因子，是治疗肺癌的新靶点。
Emerging studies suggest that microRNAs (miRNAs) play multiple roles in cancer malignancy, including proliferation and acquisition of metastatic potential. Differentially expressed miRNAs responsible for the malignancy of lung cancer were searched by miRNA microarray using a previously established brain metastatic lung cancer model. Twenty-five miRNAs were down-regulated in brain metastatic lung cancer cells. Among those, miR-193b-3p and -5p were chosen for further studies. Their function in metastatic potential and proliferation was examined using Transwell invasion, wound healing, and colony forming assays. The underlying mechanism of tumor-suppressor miR-193b-3p and -5p was explored using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR), Western blot, Argonaute 2-RNA immunoprecipitation (Ago2-RIP), and reporter assays. Both strands of miR-193b were down-regulated in brain metastatic lung cancer cells and in tissues from lung cancer patients. Overexpression of miR-193b-3p and -5p inhibited invasive and migratory activities and diminished clonogenic ability. Conversely, inhibition of miR-193b-3p or -5p increased the metastatic potential and colony forming ability. Cyclin D1 (CCND1), Ajuba LIM Protein (AJUBA), and heart development protein with EGF like domains 1 (HEG1) were identified as common target genes of miR-193b-3p and -5p. A reporter assay and an Ago2-RIP experiment showed that both miRNAs directly bind to the 3' untranslated region (3'UTR) of the target mRNA. Knockdown of target gene reduced the proliferative and metastatic potential of primary and metastatic lung cancer cells. Our results demonstrate miR-193b is a dual-strand tumor suppressor and a novel therapeutic target for lung cancer.