Department of Gastroenterology, The Xiangtan Central Hospital, Xiangtan, China.
Mol Cell Biochem. 2013 Sep;381(1-2):233-42. doi: 10.1007/s11010-013-1707-6. Epub 2013 Jun 7.
A previous study demonstrated that miR-126 expression was significantly downregulated in highly metastatic colon cancer cells. This study was to investigate the biological function of miR-126 and its regulation of target genes in colon cancer cells. Quantitative PCR was used to detect miR-126 expression in colon cancer SW480 and SW620 cells. MTT assay was to measure the changed cell viability after miR-126 mimics transfection. Wound healing and Transwell migration and invasion assays measured capacity of tumor cell migration and invasion of SW480 and SW620 cells after miR-126 transfection. Luciferase reporter assay and Western blot were used to assess both transcriptional and expression levels of one of the miR-126 target genes (i.e., CXCR4). Levels of miR-126 expression were lower in colon cancer SW480 and SW620 cells than in the adjacent normal epithelial tissues (P < 0.05). Transfection of miR-126 mimics significantly reduced colon cancer cell viability compared to NC cells (P < 0.05). The wound healing and Transwell migration and invasion assays showed that miR-126 mimics inhibited SW480 and SW620 cell migration and invasion capacity. Bioinformatics predicted that CXCR4 is one of the miR-126 target genes. Indeed, luciferase reporter assay and Western blot confirmed that CXCR4 is a miR-126 target gene. Expression of miR-126 inhibited colon cancer cell viability and reduced tumor cell migration and invasion capacity by its negative regulation of CXCR4 expression.
先前的研究表明，miR-126 的表达在高转移性结肠癌细胞中显著下调。本研究旨在探讨 miR-126 在结肠癌细胞中的生物学功能及其对靶基因的调控。定量 PCR 用于检测结肠癌细胞 SW480 和 SW620 中 miR-126 的表达。MTT 检测 miR-126 模拟物转染后细胞活力的变化。划痕愈合和 Transwell 迁移和侵袭实验测量 miR-126 转染后 SW480 和 SW620 细胞的肿瘤细胞迁移和侵袭能力。荧光素酶报告基因检测和 Western blot 用于评估 miR-126 靶基因之一（即 CXCR4）的转录和表达水平。与相邻正常上皮组织相比，结肠癌 SW480 和 SW620 细胞中的 miR-126 表达水平较低（P < 0.05）。与 NC 细胞相比，miR-126 模拟物转染显着降低结肠癌细胞活力（P < 0.05）。划痕愈合和 Transwell 迁移和侵袭实验表明，miR-126 模拟物抑制 SW480 和 SW620 细胞的迁移和侵袭能力。生物信息学预测 CXCR4 是 miR-126 的靶基因之一。事实上，荧光素酶报告基因检测和 Western blot 证实 CXCR4 是 miR-126 的靶基因。miR-126 通过负调控 CXCR4 的表达抑制结肠癌细胞活力，并降低肿瘤细胞迁移和侵袭能力。
