Effect Of RNA Interference Targeting MT1-MMP On The Expression Of Human Lung Adenocarcinoma A549Cells

Objective By gene transfection and RNAi technology, the chemical synthesis smallinterfering RNA （siRNA） specific targeted to human membrane type matrixmetalloproteinases-1(MT1-MMP), to explore whether siRNA-MT1-MMP couldeffectively transfect into human lung adenocarcinoma A549cells and inhibit theMT1-MMP gene expression in human lung adenocarcinoma A549cells.Methods Human MT1-MMP gene sequence as a template in accordance with theprinciple of siRNA design, chemical synthesis of three double-stranded siRNA, namedsiRNA-1, siRNA-2, siRNA-3. These siRNA were transiently transfected into A549cellsthrough Lipofectamine2000. Five groups were selected for the study: blank control group,negative control group and the three RNAi groups. The best mix proportion ofLipofectamine2000and siRNA was checked by flow cytometry. The transfectioneffeciency was confirmed by fluorescence microscope. While24hours and48hours after transfection, the MT1-MMP mRNA level and the expression of MT1-MMP protein wasdetected by qRT-PCR, Western blot, respectively.Results The three double-stranded siRNA could successfully transfected into lungadenocarcinoma A549cells.The best mix proportion of Lipofectamine2000and siRNAwas (1:2)and the transfection effeciency after12h was80.18%. in transfected24h,48h,respectively the collection of cells, compare blank group to interference group theexpression of MT1-MMP gene was significantly reduced, the expression of MT1-MMPin negative control and blank control was no significant difference.48h after transfectionof siRNA MT1-MMP-3, the expression of MT1-MMP mRNA and protein wasdown-regulated significantly.Conclusion The experimental use of RNAi technology,effectively down-regulates theexpression of MT1-MMP gene in human lung adenocarcinoma A549cells．To furtherexplore the MT1-MMP gene in human lung cancer migration and invasion of the importantrole of the foundation．...