【摘要】 目的观察膜型基质金属蛋白酶-1(MT1-MMP)对人肝癌细胞株浸润能力的影响,并初步探讨其作用机制。方法将稳定表达MT1-MMP基因转染至HepG2细胞中,应用Western印迹法检测MT1-MMP蛋白表达;明胶酶谱分析法检测MMP-2酶原活性;体外侵袭实验检测细胞株浸润能力。结果重组质粒转染株MT1-MMP蛋白表达水平明显高于空质粒组和未转染组(P<0.01)。明胶酶谱分析实验发现,MT1-MMP组同时检测到72000酶原形式和64000活性形式的MMP-2,另两组只检测到72000酶原形式的MMP-2。体外侵袭实验结果显示,MT1-MMP组细胞穿过基质胶(Matrigel)的细胞数目明显多于其他两组(P<0.01)。结论MT1-MMP能显著增强肝癌细胞株的浸润能力,其机制主要是通过激活MMP-2酶原,降解肿瘤周围的基质成分实现的。更多还原

【Abstract】 Objective To observe the effects of membrane type-1 matrix metalloproteinase(MT1-MMP) on the invasiveness of human hepatocellular carcinoma cells and to investigate its mechanisms.Methods HepG2 with overexpression of MT1-MMP were established by stable transfection.MT1-MMP protein was detected by Western blot.Latent pro-MMP-2 and MMP-2 activity were analyzed by gelatin zymography.The invasiveness of cell strain was measured by invasion chamber coated with Matrigel.Results MT1-MMP protein level of HepG2 cells that transfected with the recombinant vector was obviously higher than that of empty vector group and that of control group(P<0.01).There were both 72 000 precursor form and 64 000 active enzyme form of MMP-2 in group that transfected with the recombinant vector,but there was only 72 000 precursor form of MMP-2 in empty vector group and control group.The largest amount of cells penetrated through Matrigel was observed in group that transfected with the recombinant vector than that in the other two groups(P<0.01).Conclusions MT1-MMP can remarkably promote the invasive potential of hepatocellular carcinoma cells mainly through its ability of activating latent pro-MMP2 to degrade extracellular matrix.更多还原