过表达P185可抑制EMT并促进胃癌细胞侵袭和迁移

Overexpression of P185 might Promote Invasion and Migration of Gastric Cancer Cells by Inhibiting EMT

为探究过表达P185基因对胃癌SGC7901细胞侵袭、迁移的影响以及可能作用机制,本研究通过脂质体将携带P185基因的过表达pcDNA3.1-P185质粒,转染至胃癌SGC7901细胞中;本研究采用qRT-PCR和免疫印迹试验(Western blotting)检测P185 mRNA的转录水平和蛋白水平,Western blotting检测钙黏蛋白E(E-cadherin)、波形蛋白(Vimentin)、基质金属蛋白酶-2 (matrix metalloprotease, MMP-2)表达;以Transwell小室法检测细胞的侵袭、迁移能力的变化。研究结果表明:胃癌细胞转染过表达pcDNA3.1-P185质粒能显著上调P185 mRNA和蛋白质的表达(p<0.05);SGC7901细胞转染重组质粒pcDNA3.1-P185后,细胞的侵袭、迁移能力较对照组显著增强(p<0.05),细胞中E-cadherin蛋白水平显著下调(p<0.05),Vimentin、MMP-2蛋白水平显著增加(p<0.05)。本研究显示P185可能通过抑制EMT,促进细胞外基质的降解、促进胃癌细胞的侵袭、迁移。

To investigate the effect of overexpression P185 gene on invasion and migration of gastric cancer SGC7901 cells and its possible mechanism, the overexpression vector pcDNA3.1-P185 carrying P185 gene was transfected into gastric cancer SGC7901 cells by LipofectamineTM2000. The expression of P185 was detected by q RT-PCR and Western blotting. Western blotting was used to detect the expression of E-cadherin, vimentin, MMP-2;Transwell methods were used to detect the changes of cell invasion, migration. The results indicated that overexpression of pcDNA3.1-P185 plasmid could up-regulate the expression of P185 mRNA and protein in gastric cancer cells(p<0.05), the ability of cell invasion and migration were significantly higher than those of the control group(p<0.05). The level of E-cadherin protein was significantly down-regulated(p<0.05), but Vimentin,MMP-2 protein increased significantly(p<0.05). This study showed that P185 might promote the degradation of extracell-ular matrix as well as the invasion and migration of gastric cancer cells by inhibiting EMT.