Du Ying, Wang Lingfei, Wu Honghai, Zhang Yiyin, Wang Kan, Wu Dingting
Department of Endocrinology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, P.R. China.
Department of Pharmacy, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310009, P.R. China.
Mol Med Rep. 2015 Sep;12(3):3416-3422. doi: 10.3892/mmr.2015.3789. Epub 2015 May 15.
Human microRNA (miR)-141 is a member of the miR‑200 family, which has been reported to be downregulated in gastric cancer, and involved in the proliferation of gastric cancer cells. However, little is currently known regarding its role in the migration of gastric cancer. The present study investigated the function of miR‑141 in gastric cancer cell migration, and evaluated the contribution of zinc finger E‑box‑binding homeobox 1 and 2 (ZEB1/2) in miR‑141 mediated migration of gastric cancer cells. The expression levels of miR‑141 and its potential ZEB1/2 targets were examined by quantitative polymerase chain reaction (qPCR) and western blotting, respectively. The migration of SGC‑7901 and HGC‑27 gastric cancer cells, which had been transfected with an miRNA precursor, was examined by cell migration and wound healing assays. A luciferase activity assay was used to validate whether ZEB1/2 was a direct target of miR‑141. The results demonstrated that overexpression of miR‑141 markedly inhibited the migration of gastric cancer cells in vitro. Forced overexpression of miR‑141 significantly reduced the luciferase activity of the 3'‑untranslated region of ZEB2 in gastric cancer cells. Furthermore, the mRNA and protein expression levels of ZEB2 were reduced in cells overexpressing miR‑141, whereas the protein expression levels of E‑cadherin were increased. In gastric tumor samples the expression levels of ZEB2 were inversely correlated with the expression of miR‑141. These results suggest that miR‑141 may be involved in the inhibition of gastric cancer cell migration, and that ZEB2 is a target gene of miR-141.
人微小RNA(miR)-141是miR-200家族的成员,据报道其在胃癌中表达下调,并参与胃癌细胞的增殖。然而,目前关于其在胃癌迁移中的作用知之甚少。本研究调查了miR-141在胃癌细胞迁移中的功能,并评估了锌指E盒结合同源框1和2(ZEB1/2)在miR-141介导的胃癌细胞迁移中的作用。分别通过定量聚合酶链反应(qPCR)和蛋白质印迹法检测miR-141及其潜在的ZEB1/2靶标的表达水平。通过细胞迁移和伤口愈合试验检测转染了miRNA前体的SGC-7901和HGC-27胃癌细胞的迁移情况。使用荧光素酶活性测定法验证ZEB1/2是否为miR-141的直接靶标。结果表明,miR-141的过表达显著抑制了体外胃癌细胞的迁移。miR-141的强制过表达显著降低了胃癌细胞中ZEB2 3'非翻译区的荧光素酶活性。此外,在过表达miR-141的细胞中,ZEB2的mRNA和蛋白质表达水平降低,而E-钙黏蛋白的蛋白质表达水平升高。在胃肿瘤样本中,ZEB2的表达水平与miR-141的表达呈负相关。这些结果表明,miR-141可能参与抑制胃癌细胞迁移,并且ZEB2是miR-141的靶基因。