Objective:To investigate the activation and mechanism of MMP-2.Methods:Examined the expression of MMP-2 by zymography,and the expression of MT1-MMP mRNA were examined by reverse transcription polymerase chain reaction (RT-PCR).Matrigel invasion of MCF-7 cells was detected with boyden chamber.Results:Induction of MMP-2 activation was specific to MCF-7 cells cultured on type I collagen gel.MT1-MMP mRNA was expressed in MCF-7 cells,and enhanced expression of MT1-MMP mRNA was seen in cells cultured on type I collagen gel.Treated with the antibody MT1-MMP inhibits the cell surface gelatinolytic activity,Matrigel invasion of MCF-7 cells by treated with the antibody MT1-MMP was reduced.Conclusion:Culture of MCF-7 cells on 3D-type I collagen gel induced MMP-2 activation,suggesting that the activation of MMP-2 was triggered by an interaction between MCF-7 cells and ECM components,MMP-2 actived by MT1-MMP promotes the matrigel invasion of MCF-7 cells.%目的:研究MMP-2的活化并探索其机制.方法:采用酶谱分析方法检测瘤细胞条件培养基中MMP-2在包被前后表达和活化的情况,用RT-PCR方法检测MT1-MMP mRNA的表达,观察MT1-MMP激活的MMP-2在细胞侵袭中的作用,Boyden小室膜侵袭实验检测细胞的侵袭能力.结果:条件培养基明胶酶谱分析结果显示仅在三维聚I型胶原包被组存在MMP-2的活性形式;培养于I型胶原组的人MCF-7细胞MT1-MMP mRNA表达量明显高于对照组,与对照组间差异显著(P<0.01).Boyden小室膜侵袭实验可见,用MT1-MMP的抗体处理组的细胞数明显少于未用MT1-MMP的抗体处理组,差异有统计学意义(P<0.01).结论:I型胶原成分可以通过上调MT1-MMP的水平来调控人乳腺癌MCF-7细胞MMP-2的活化,MT1-MMP激活的MMP-2可以增强人乳腺癌MCF-7细胞的侵袭能力.
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