miR-874 Inhibits cell proliferation, migration and invasion through targeting aquaporin-3 in gastric cancer
miR-874 Inhibits cell proliferation, migration and invasion through targeting aquaporin-3 in gastric cancer
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DOI：
10.1007/s00535-013-0851-9
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发表时间：
2014-06-01
影响因子：
6.3
通讯作者：
Xu, Zekuan
中科院分区：
文献类型：
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作者：
Jiang, Baofei;Li, Zengliang;Xu, Zekuan
水通道蛋白3（Aquaporin-3，AQP 3）是一种水转运蛋白，在多种恶性肿瘤中起致瘤作用。然而，其监管机制迄今仍不明确。本研究通过生物信息学方法寻找靶向AQP 3的microRNA，并通过荧光素酶报告基因分析、microRNA RT-PCR和western blotting等方法对microRNA进行鉴定，以探讨microRNA介导的AQP 3基因表达抑制机制。采用microRNA RT-PCR和Western blotting方法检测miR-874和AQP 3在人胃癌标本和胃癌细胞株中的表达情况。采用5-乙炔基-2 '-脱氧尿苷、细胞迁移和侵袭实验以及体内致瘤实验，观察miR-874缺失或异位表达对胃癌细胞表型的影响。流式细胞术检测胃癌细胞凋亡情况，TUNEL法检测胃癌细胞凋亡情况，miR-874通过与胃癌细胞AQP 3 mRNA的3 'UTR结合抑制胃癌细胞AQP 3的表达。miR-874在临床样本中显著下调，并且与AQP 3蛋白水平显著相关。临床病理学意义分析显示miR-874和AQP 3与胃癌特征密切相关。功能分析表明，异位表达miR-874抑制GC细胞的生长、迁移、侵袭和致瘤性，而敲低miR-874则促进这些表型。miR-874通过下调Bcl-2、MT 1-MMP、MMP-2和MMP-9的表达，上调caspase-3和Bax的活性，诱导细胞凋亡，抑制细胞迁移和侵袭，为miR-874上调AQP 3的表达提供了可能的机制，同时也表明miR-874在胃癌发生发展中的重要作用。
Aquaporin-3 (AQP3) is a water transporting protein which plays an oncogenic role in several malignant tumors. However, its regulatory mechanism remains elusive to date. In this study, we investigated the microRNA-mediated gene repression mechanism involved in AQP3's role.The potential microRNAs targeting AQP3 were searched via bioinformatic methods and identified by luciferase reporter assays, microRNA RT-PCR and western blotting. The expression patterns of miR-874 and AQP3 in human gastric cancer (GC) specimens and cell lines were determined by microRNA RT-PCR and western blotting. 5-ethynyl-2'-deoxyuridine, cell migration and invasion assays and tumorigenicity in vivo were adopted to observe the effects of miR-874 depletion or ectopic miR-874 expression on GC cell phenotypes. Cell apoptosis was evaluated by FACS and TUNEL in vitro and in vivo respectively.miR-874 suppressed AQP3 expression by binding to the 3'UTR of AQP3 mRNA in GC cells. miR-874 was significantly down-regulated and reversely correlated with AQP3 protein levels in clinical samples. Analysis of the clinicopathological significance showed that miR-874 and AQP3 were closely correlated with GC characteristics. Functional analyses indicated that ectopic miR-874 expression suppressed the growth, migration, invasion and tumorigenicity of GC cells, whereas miR-874 knockdown promoted these phenotypes. Down-regulation of Bcl-2, MT1-MMP, MMP-2 and MMP-9 and upregulation of caspase-3 activity and Bax were involved in miR-874 inducing cell apoptosis, and inhibiting migration and invasion.These results provide a mechanism by which AQP3 is upregulated, as well as highlight the importance of miR-874 in gastric cancer development and progression.