miR-582-5p通过诱导EMT抑制胃癌细胞的侵袭转移

The miR-582-5p inhibits invasion and migration of gastric cancer cells by inducing epithelial-mesenchymal transition

目的 探讨miR-582-5p对胃癌细胞侵袭迁移的影响,并初步探讨其分子机制。方法 使用miR-582-5p mimics转染胃癌SGC-7901和AGS细胞,real-time PCR验证转染率。后续实验分为实验组(转染miR-582-5p mimics)和空白对照组(转染negative control),Transwell和细胞划痕实验检测上调miR-582-5p对SGC-7901和AGS细胞侵袭、迁移的作用;real-time PCR和Western blotting检测上调miR-582-5p后对上皮-间质转化(epithelial-mesenchymal transition,EMT)基因Slug、Snail、vimentin、fibronectin和E-cadherin的影响。Dickkopf-1阻断Wnt/β-catenin信号通路后观察上调miR-582-5p对SGC-7901和AGS细胞侵袭的影响。结果 转染miR-582-5p mimics的SGC-7901和AGS细胞miR-582-5p的表达较胃黏膜细胞表达显著降低(均 P <0.05)。上调miR-582-5p后SGC-7901和AGS细胞的侵袭、迁移能力显著降低;上调miR-582-5p后SGC-7901和AGS细胞中Slug、Snail、vimentin及fibronectin蛋白的表达显著降低,E-cadherin表达显著增高(均 P <0.05)。Dickkopf-1减弱了上调miR-582-5p对SGC-7901和AGS细胞的侵袭的抑制作用。结论 胃癌细胞上调miR-582-5p后可能通过诱导EMT降低细胞侵袭、迁移能力,这可能和Wnt/β-catenin激活有关。

Objective To investigate the influence of miR-582-5p on invasion and migration of gastric cancer cells and its underlying mechanism. Methods The expression of miR-582-5p in gastric cancer cells was detected by real-time PCR. SGC-7901 and AGS cells were transfected with miR-582-5p mimics, and miR-582-5p expression was determined by real-time PCR. Transwell and would healing assays were employed to detect the effect of miR-582-5p up-regulation on invasion and migration. Levels of Slug, Snail, vimentin, fibronectin and E-cadherin were detected after miR-582-5p up-regulation by Western blot analysis and real-time PCR. The effect of blockage of the Wnt/β-catenin pathway by Dickkopf-1 on invasion of SGC-7901 and AGS cells was investigated. Results The transfection of miR-582-5p mimics up-regulated miR-582-5p expression in SGC-7901 and AGS cells(all P <0.05). In SGC-7901 and AGS cells after transfection with miR-582-5p mimics, the invasive and migratory abilities of gastric cancer cells were inhibited( P <0.05). The protein levels of Slug, Snail, vimentin and fibronectin were decreased, while E-cadherin was increased after up-regulation of miR-582-5p(all P <0.05). Dickkopf-1 treatment resulted in a significant inhibition in invasive abilities of SGC-7901 and AGS cells decreased by up-regulation of miR-582-5p(all P <0.05). Conclusion The up-regulation of miR-582-5p may inhibit invasion and migration of gastric cancer cells by inducing epithelial-mesenchymal transition, which is related to the activation of Wnt/β-catenin pathway.