[目的]构建基质金属蛋白酶3(Matrix metalloproteinase3,MMP3)基因过表达慢病毒载体,修饰胚胎干细胞(ES)检测其对成纤维细胞的作用。[方法]克隆MMP3基因,构建EGFP标记的MMP3慢病毒表达载体,用GP2-293T细胞包装慢病毒,修饰ES细胞,与成纤维细胞3T3共培养。镜下观察绿色荧光蛋白表达,Western Blot检测MMP3和TGF-β蛋白表达。[结果]成功钓取MMP3基因,并构建慢病毒表达载体LV-MMP3-EGFP;在转染的GP2-293T细胞中表达绿色荧光蛋白;修饰后的ES细胞表达绿色荧光蛋白,72 h蛋白表达量最高,转染效率达到80%以上;被修饰细胞中MMP3蛋白表达高于对照组,其表达量是对照组的1.5倍;共培养后3T3细胞中TGF-β蛋白表达实验组与对照组无显著变化。[结论]LV-MMP3-EGFP修饰的ES细胞能够大量表达MMP3蛋白,作用于细胞外基质,但是对成纤维细胞中TGF-β表达没有显著影响(P>0.05)。 [Objective] To construct a lentiviral vector overexpression matrix metalloproteinase 3 (MMP3) gene and modify embryonic stem cells (ES) to detect its effect on fibroblasts. [Method] The MMP3 gene was cloned and EGFP-labeled MMP3 lentiviral vector was constructed. GP2-293T cells were packaged with lentivirus, ES cells were modified and co-cultured with fibroblast 3T3. The green fluorescence protein expression was observed microscopically. The protein expressions of MMP3 and TGF-β were detected by Western Blot. [Results] The gene of MMP3 was successfully captured and the lentiviral expression vector LV-MMP3-EGFP was constructed. The green fluorescent protein was expressed in the transfected GP2-293T cells. The green fluorescent protein was expressed in the modified ES cells, The highest transfection efficiency reached more than 80%; modified cells MMP3 protein expression was higher than the control group, the expression of 1.5 times that of the control group; 3T3 cells co-cultured TGF-βprotein expression in the experimental group and the control group no significant Variety. [Conclusion] LV-MMP3-EGFP-modified ES cells can express MMP3 protein in large quantities and act on the extracellular matrix, but have no significant effect on the expression of TGF-β in fibroblasts (P> 0.05).