RNAi沉默中期因子基因表达对胃癌细胞黏附和侵袭力的影响

Effects of RNA interference midkine gene expression on cell adhesion and invasion of human gastric cancer cell

目的:探讨中期因子(midkine,MK)基因小干扰RNA对胃癌细胞生物学行为的影响。方法:培养人胃癌BGC-823、HGC-27、MGC-803及SGC-7901细胞株,以荧光实时定量PCR方法检测中期因子基因mRNA表达;筛选出中期因子表达最高者。采用中期因子基因小干扰RNA转染胃癌细胞株,分别以荧光实时定量PCR和免疫荧光方法观察中期因子基因mRNA和蛋白水平,然后以四唑蓝(MTT)比色法检测细胞黏附性,以Boyden方法检测癌细胞侵袭力。结果:4株胃癌细胞中,中期因子均有不同程度的表达,以胃癌BGC-823细胞最高;以MK siRNA转染胃癌BGC-823细胞后,癌细胞中期因子基因mRNA和蛋白水平明显下降,且呈浓度依赖性;转染组细胞黏附数量明显下降,细胞侵袭力明显下降。结论:中期因子基因在胃癌细胞黏附和侵袭中发挥着重要作用;以siRNA转染胃癌细胞,可抑制胃癌细胞黏附和侵袭能力。

Objective： To study the effects of midkine（MK） gene small interfering RNA（siRNA）on adhesion and invasion of human gastric cancer cell.Methods： Real time PCR was used to evaluate the MK mRNA expression of human gastric cancer cell lines BGC-823,HGC-27,MGC-803 and SGC-7901.The cell of MK highest expression was transfected with different dose of MK siRNA.The expression of MK mRNA and proteinwere were determined by real-time quantitative PCR and immumoflurescence method.The cell adhesion was evaluated by MTT assay,and invasion was exmined by Boyden chamber,respectively.Results： Cell line BGC-823 showed the highest elevation of MK mRNA in four gastric cancer cell lines.The results from real-time quantitative PCR and immumoflurescence method showed that MK mRNA and protein reduced in time-and dose-dependent manners（P0.01;P0.01）.The adhesive,and invasive ability of BGC-823 cell treated with MK siRNA decreased compared with control groups（P0.01,P0.01）.Conclusion： MK gene might play an important role in adhesion and invasion of human gastric cancer cell.siRNA targeted MK could effectively inhibit adhesion,migration,and invasion of human gastric cancer cell.