微小RNA-30e调控EMT对胃癌侵袭和迁移影响的研究

Effects of microRNA-30e Regulating Epithelial Mesenchymal Transition on Invasion and Migration in Gastric Cancer

目的:探讨微小RNA-30e(miR-30e)对胃癌细胞迁移和侵袭能力的影响及可能的作用机制。方法:利用Transwell实验和细胞划痕实验检测胃癌细胞系BGC823侵袭和迁移的能力;以脂质体包裹合成miR-30e转染至BGC823细胞,并设空白载体作为对照组;Real-time PCR分别检测实验组和对照组细胞中miR-30e的表达。RT-PCR检测过表达miR-30e后对上皮细胞间充质转化(EMT)相关标记分子Snail、Vimentin、N-cadherin和E-cadherin表达的影响。结果:miR-30e转染至胃癌细胞后,抑制EMT通路主要因子Snail,Vimentin和N-cadherin m RNA和蛋白质表达,而增加E-cadherin的mRNA和蛋白质表达;miR-30e通过TGF-β对BGC823细胞的侵袭和迁移能力有明显的抑制作用。结论:miR-30e可能是肿瘤细胞EMT过程的关键靶标靶点,阻断EMT过程,可以抑制胃癌细胞的侵袭和迁移能力。

Objective: To investigate the effect of microRNA-30e(miR-30e) on the invasion and migration of gastric cancer and the potential mechanism. Methods: The transwell system and the assays of wound healing were used to assess the invasion and migration ability of gastric cell line BGC823. The miR-30e was transfected into BGC823 cell via lipofectamine 2000. Cells were also transfected with empty vectors to serve as controls. The expression of miR-30e was detected by real-time PCR, and western blot was used to assay the expression of Snail, Vimentin, N-cadherin, and E-cadherin which were the markers of epithelial-mesenchymal transition(EMT).Results: After the transfection of the miR-30e, which had significantly suppressed the major factors in EMT signaling pathway, such as the expression of m RNA and proteins of Snail, N-cadherin, and vimentin, increased the m RNA and proteins expression of E-cadherin.And the mimic also had suppressed the invasion and migration of BGC823 cell line induced by TGF-β. Conclusion: MiR-30e may be a key target in the EMT process of cancer cells, blocking the EMT process could inhibit the invasion and migration of gastric cancer cells.