miR-375 inhibits the invasion and metastasis of colorectal cancer via targeting SP1 and regulating EMT-associated genes
miR-375 inhibits the invasion and metastasis of colorectal cancer via targeting SP1 and regulating EMT-associated genes
复制标题
DOI：
10.3892/or.2016.4834
复制
发表时间：
2016-07-01
期刊：
影响因子：
4.2
通讯作者：
Zhu, Hongguang
中科院分区：
文献类型：
--
作者：
Cui, Fengyun;Wang, Shuyang;Zhu, Hongguang
越来越多的证据表明，异常表达的microRNAs （miRNAs）与肿瘤的发生和进展有关。我们之前的研究发现microRNA-375 （miR-375）在结直肠癌（CRC）中下调，但对miR-375在结直肠癌发生中的作用和相关机制知之甚少。通过细胞计数试剂盒-8 （CCK-8）、菌落形成和加或不加Matrigel的Transwell实验研究了细胞的增殖、侵袭和迁移效应。此外，筛选候选靶基因，并通过荧光素酶报告基因和western blot检测进行验证。此外，通过western blot分析探讨上皮-间质转化（epithelial-mesenchymal transition， EMT）的分子机制。发现miR-375抑制DLD1和HCT8细胞的增殖、侵袭和迁移。此外，miR-375通过直接结合3'-非翻译区（3'-UTR）负向调节Spl转录因子（SP1）蛋白。此外，我们发现miR-375调控基质金属蛋白酶2 （MMP2）和emt相关基因E-cadherin、vimentin、snail、N-cadherin和β -catenin。总之，miR-375通过直接靶向SP1，调控MMP2和emt相关基因，抑制了细胞的增殖、侵袭和迁移。
Accumulating evidence has shown that aberrantly expressed microRNAs (miRNAs) are associated with tumor development and progression. Our previous study found that microRNA-375 (miR-375) was downregulated in colorectal cancer (CRC), but little is known concerning the role of miR-375 and the related mechanism in CRC development. The proliferation, invasion and migration effects were investigated by Cell Counting Kit-8 (CCK-8), colony formation and Transwell assays with or without Matrigel. In addition, candidate target genes were screened and validated by luciferase reporter and western blot assays. In addition, western blot analysis was performed to explore the molecular mechanisms associated with epithelial-mesenchymal transition (EMT). It was found that miR-375 inhibited proliferation, invasion and migration in DLD1 and HCT8 cells. In addition, miR-375 negatively regulated Spl transcription factor (SP1) protein by directly binding to the 3'-untranslated region (3'-UTR). Furthermore, it was found that miR-375 regulated matrix metalloproteinase 2 (MMP2) and EMT-associated genes, E-cadherin, vimentin, snail, N-cadherin and beta-catenin. In conclusion, miR-375 inhibited the proliferation, invasion and migration by directly targeting SP1 and regulating MMP2 and EMT-associated genes.