膜型基质金属蛋白酶-1在基质金属蛋白酶-2活化中的作用机制

Involvement of membrane type 1-matrix metalloproteinase in the activation ofmatrix metalloproteinase-2

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。

Objective To investigate the activation and mechanism of MMP -2. Methods ： Examined the expres-sion 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. Re-sults ： Induction of MMP -2 activation was specific to MCF -7 cells cultured on type I collagen gel. MT1 - MMP mR- NA 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 inva-sion 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 in-teraction between MCF -7 cells and ECM components,MMP -2 actived by MT1 - MMP promotes the matrigel inva-sion of MCF - 7 cells.