摘要
目的研究AKT1基因沉默对人喉癌细胞系Hep-2体外侵袭能力的影响。方法利用阳离子脂质体将AKT1siRNA转染Hep-2细胞,RT-PCR和Western blot分析各组细胞AKT1,MMP-2mRNA和蛋白表达,划痕实验观察细胞迁移能力,Transwell小室细胞测定细胞侵袭能力。结果 AKT1siRNA可抑制AKT1mRNA转录和蛋白表达水平;AKT1基因沉默后细胞迁移运动能力明显减弱,穿模细胞数明显减少(P<0.01);MMP-2mRNA和蛋白表达下调。结论 AKT1siRNA可通过下调MMP-2表达抑制Hep-2细胞体外侵袭能力。
Objective To investigate the influence of AKT1 gene silencing on the invasion ability of Hep-2 cell . MethoO. s siRNA targeting AKT1 was transfected into Hep-2 cell by lipofectamine. RT-PCR and western blot tech- nique were used to test the the expression of AKT1 ,MMP-2 mRNA and protein. Scratch assay and transwell chamber experiment were used to analyze the invasion and migration ability. Results The mRNA and protein expression level of AKT1 gene was inhibited by AKTlsiRNA. The ability of migration and invasion declined obviously after AKTlgene si- lencing,number of cells invading through membrane was decreased. The mRNA and protein expression level of MMP-2was downregulated. Conclusion By down-regulating the expression of MMP-2,the invasion ability of Hep 2 cells was inhibited with AKTlsiRNA.
出处
《中国实验诊断学》
2013年第11期1959-1963,共5页
Chinese Journal of Laboratory Diagnosis
基金
辽宁省博士启动基金资助项目(20081042)