Zhang Hengchun, Sun Zhiguo, Li Yan, Fan Dong, Jiang Hao
Department of General Surgery, Hongqi Hospital of Mudanjiang Medical University, Mudanjiang, Heilongjiang, 157011, China.
Department of General Surgery, Hongqi Hospital of Mudanjiang Medical University, Mudanjiang, Heilongjiang, 157011, China.
Biomed Pharmacother. 2017 Apr;88:285-292. doi: 10.1016/j.biopha.2017.01.023. Epub 2017 Jan 20.
We aimed to investigate the effects of miR-200c binding to fibronectin 1 (FN1) on proliferation, migration and invasion of gastric cancer (GC) cells. A total of 52 GC tissues and their corresponding normal adjacent tissue samples were collected. Then, miR-200c and FN1 were tested using quantitative real-time RT-PCR in the clinical specimens and GC cells, while immunohistochemistry and western blotting assay were carried out to detect FN1 expressions. Dual luciferase reporter gene assay was used to assess the effect of miR-200c on the luciferase activity of FN1 3'UTR. BGC-823 cells were transfected with miR-200c mimics, miR-200c inhibitors and FN1 siRNA, respectively. The effects of miR-200c inhibitors and FN1 siRNA on cellular proliferation, migration and invasion were detected through MTT assay and Transwell assay. Compared to normal tissues and cells, miR-200c was significantly down-regulated and FN1 was significantly up-regulated (P<0.01). Dual luciferase reporter gene assay showed that miR-200c could specifically bind to the 3'-UTR of FN1 and significantly repress the luciferase activity (P<0.01). Both mRNA and protein expressions of FN1 were decreased significantly in GC cells when miR-200c was over expressed. The proliferation, migration and invasion of GC cells could be suppressed by over-expression of miR-200c or down-regulation of FN1. In conclusion, miR-200c was significantly down-regulated in both GC tissues and cell lines, while FN1 presented the opposite trends. Besides, miR-200c inhibited the proliferation, migration and invasion of GC cells through binding to FN1.
我们旨在研究微小RNA-200c（miR-200c）与纤连蛋白1（FN1）结合对胃癌（GC）细胞增殖、迁移和侵袭的影响。共收集了52例GC组织及其相应的正常相邻组织样本。然后，采用定量实时逆转录聚合酶链反应（RT-PCR）检测临床标本和GC细胞中的miR-200c和FN1，同时进行免疫组织化学和蛋白质免疫印迹分析以检测FN1的表达。采用双荧光素酶报告基因测定法评估miR-200c对FN1 3'非翻译区（UTR）荧光素酶活性的影响。分别用miR-200c模拟物、miR-200c抑制剂和FN1小干扰RNA（siRNA）转染BGC-823细胞。通过MTT法和Transwell法检测miR-200c抑制剂和FN1 siRNA对细胞增殖、迁移和侵袭的影响。与正常组织和细胞相比，miR-200c显著下调，FN1显著上调（P<0.01）。双荧光素酶报告基因测定法显示，miR-200c可特异性结合FN1的3'-UTR并显著抑制荧光素酶活性（P<0.01）。当miR-200c过表达时，GC细胞中FN1的mRNA和蛋白表达均显著降低。miR-200c的过表达或FN1的下调均可抑制GC细胞的增殖、迁移和侵袭。总之，miR-200c在GC组织和细胞系中均显著下调，而FN1呈现相反的趋势。此外，miR-200c通过与FN1结合抑制GC细胞的增殖、迁移和侵袭。
