小鼠基质金属蛋白酶-9 RNA干扰慢病毒载体在体外的沉默效应

The silencing effect of MMP-9 RNA interfere lentivirus vector of the mice in vitro

目的探讨构建的小鼠基质金属蛋白酶(MMP)-9 RNA干扰(RNAi)慢病毒载体在体外工具细胞中的沉默效应。方法针对小鼠MMP-9基因序列构建的MMP-9过表达克隆质粒和RNAi慢病毒载体共转染培养良好的工具细胞(293T和NIH3T3细胞),采用实时荧光定量PCR和Western印迹观察RNAi对工具细胞中MMP-9 mRNA和蛋白表达的下调作用。结果构建的MMP-9 RNAi慢病毒载体能显著降低工具细胞中MMP-9表达量。结论成功构建了能够抑制小鼠MMP-9表达的RNAi慢病毒载体,为在体研究奠定了基础。

Objective To explore the silencing effect of the constructed mice′s matrix metalloproteinase （MMP）-9 RNA interference （RNAi） lentivirus vector in vitro. Methods 293T and NIH3T3 cells were cocultured by MMP-9 overexpressed clone plasmid and RNAi lentivirus vector. Real time PCR and Western blot were adopted to observe the downregulation of MMP-9 mRNA expression in 293T and NIH3T3 cells by RNAi. Results The constructed MMP-9 RNAi lentivirus vector significantly decreased MMP-9 expression in 293T and NIH3T3 cells. Conclusions RNAi lentivirus vector inhibiting MMP-9 expression is successfully constructed,laying foundation for the research in vivo.