Regulatory Effects of miR-205 on Invasion and Migration of Gastric Cancer Cells and Potential Mechanism
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摘要:目的
探讨miR-205对胃癌HGC27细胞侵袭、迁移的作用及其机制。
方法实时荧光定量PCR法测定4种不同胃癌细胞（AGS、MKN74、HGC27、SGC7901）中miR-205的表达情况。体外培养的HGC27细胞通过转染miR-205 mimic上调细胞中miR-205的表达，划痕实验和Transwell实验观察其侵袭和迁移能力；Western blot检测上皮间质转化（epithelial-mesenchymal transition, EMT）进程及相关信号通路的活性。
结果miR-205在4种胃癌细胞中均呈低表达（均P < 0.05）。过表达miR-205后，HGC27细胞的迁移率由（54.7±4.1）%降低至（34.1±4.5）%（P=0.005）；细胞的侵袭率由（52.8±6.3）%降低至（32.2±4.9）%（P=0.001）。此外，过表达miR-205之后，胃癌细胞中上皮细胞标志物E-cadherin、β-catenin蛋白表达显著上升（P=0.002, P=0.003），而间质细胞标志物N-cadherin、vimentin的蛋白表达显著下降（P=0.005, P=0.004），Notch和snail的蛋白表达水平也显著降低（P=0.002, P=0.003）。
结论miR-205在胃癌细胞中低表达，且与胃癌细胞的侵袭和迁移密切相关；其分子机制可能与抑制EMT及Notch/snail信号通路有关。
Abstract:ObjectiveTo investigate the effect of miR-205 on the invasion and migration of HGC27 cells and related mechanism.
MethodsThe expression of miR-205 in four kinds of gastric cancer cell lines (AGS, MKN74, HGC27, SGC7901) were measured by real-time quantitative PCR. miR-205 mimic was transfected into HGC27 cells for up-regulation of miR-205, then cells invasion and migration were detected by scratch test and Transwell assay. Furthermore, epithelial to mesenchymal transition (EMT) progress and the activation of related signal pathway were measured by Western blot.
ResultsReal-time quantitative PCR showed that miR-205 was significantly down-regulated in gastric cancer cells (all P < 0.05). After miR-205 mimic transfection, miR-205 was overexpressed; the relative cellular migration rate of HGC27 cells decreased from (54.7±4.1)% to (34.1±4.5)% (P=0.005); and the cellular invasion rate decreased from (52.8±6.3)% to (32.2±4.9)% (P=0.001). Furthermore, after miR-205 was over-expressed, the expression of E-cadherin, β-catenin were up-regulated (P=0.002, P=0.003) and the expression of N-cadherin, vimentin were down-regulated (P=0.005, P=0.004) and the protein expression of Notch and snail were down-regulated (P=0.002, P=0.003).
ConclusionmiR-205 is down-regulated in gastric cancer cells and it is closely related to the invasion and migration of HGC27 cells. The molecular mechanism maybe related to the inhibition of EMT progress and Notch/snail pathway.
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Key words:
- miR-205 /
- EMT /
- Gastric cancer /
- Invasion /
- Migration
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图 1 qRT-PCR检测胃癌细胞中miR-205的表达水平以及miR-205 mimic的转染效率
Figure 1 miR-205 expression in different gastric cancer cells and transfection efficacy of miR-205 mimic detected by qRT-PCR assay
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图 2 划痕实验检测过表达miR-205对HGC27细胞迁移的影响
Figure 2 Effect of miR-205 overexpression on HGC27 cells migration detected by scratch test
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图 3 Transwell实验检测过表达miR-205对HGC27细胞侵袭的影响
Figure 3 Effect of miR-205 overexpression on HGC27 cells invasion detected by Transwell assay
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