siRNA靶向干扰COX-2基因对胃癌BGC823细胞侵袭、迁移的影响

Small Interfering RNA- mediated COX-2 Gene Silencing Inhibits the Invasion and Migration of Gastric Cancer Cell Line BGC823

目的探讨siRNA靶向干扰COX-2基因对胃癌细胞BGC823侵袭迁移能力的影响及可能机制。方法应用Lipofectamine 2000将COX-2 siRNA(siRNA-COX-2组)阴性对照siRNA(siRNA-control组)转入到细胞BGC823中,应用Transwell小室分析胃癌细胞BGC823转染前后对胃癌细胞侵袭、迁移能力的变化。应用RT-PCR检测胃癌细胞转染前后COX-2、β-catenin的mRNA水平,应用Western blot法检测胃癌细胞转染前后COX-2、β-catenin、MMP-2、MMP-9的蛋白的表达水平变化。结果 siRNA-COX-2组穿过Transwell小室微孔膜的细胞数明显下降,COX-2、β-catenin的mRNA水平及COX-2、β-catenin、MMP-2、MMP-9的蛋白的表达水平明显减少(P<0.05)。结论 COX-2基因的表达与胃癌细胞BGC823的侵袭、迁移密切相关,可能与通过抑制WNT/β-catenin信号通路抑制MMP2、MMP9有关。

Objective To investigate the effect of small interfering RNA-mediated COX-2 gene silencing inhibits on invasion and migration of gastric cancer cell line BGC823and its mechanism.Methods COX-2 siRNA was transfected into cell line BGC823 using Lipofectamine 2000.The cell migration and proliferation was evaluated using Transwell.The transcription of COX-2 and β-catenin was detected by semi-quantitative reverse transcription-polymerase chain reaction,and the protein expression level of COX-2,β-catenin,M MP-2,MMP-9 was detected by Western blot.Results The cells numbers of siRNA-COX-2 through the Transwell chambers microporous membrane was significantly decreased.The transcription of COX-2,β-catenin and the protein expression of COX-2,β-catenin,MMP-2,MMP-9 were significantly decreased.Conclusion The COX-2 gene expression is closely related to the cell invasion and migration of gastric cell line BGC823,maybe inhibit the expression of MMP2 and MMP9 by inhibit the WNT/β-catenin signaling pathway.