正义MMP-9可调控型表达与人黑色素瘤细胞系转移表型相关性的研究

目的：探讨金属蛋白酶MMP-9与肿瘤恶性表型的相关性。方法：利用基因重组技术构建正义MMP-9cDNA四环素可调控型表达载体，用脂质体法转染正义MMP-9至早期人黑色素瘤细胞株WM35(不表达MMP-9)。检测转染后细胞MMP-9表达水平的改变以及体外生长、侵袭能力的变化。结果：转染正义基因后，上调了WM35细胞MMP-9的表达及活性，细胞生长速度加快、锚着非依赖性生长能力增加、体外侵袭能力增强。而运用四环素可以抑制四环素负调控逆转录病毒载体上的外源基因的表达，结果与对照细胞相似。结论：正义MMP-9基因能促进人黑色素瘤细胞的生长和侵袭，说明MMP-9在人黑色素瘤细胞转移过程中起重要作用。同时，四环素负调控逆转录病毒载体可以调控外源基因的表达。

A STUDY ON THE CORRELATION BETWEEN THE REGULATED EXPRESSION OF MATRIX METALLOPROTEINASE 9(MMP-9) AND THE METASTATIC PHENOTYPE IN HUMAN MELANOMA CELLS

Objective To investigate the correlation between MMP-9 and tumor metastasis.Methods One self-contained tetracycline-regulated retroviral vector containing sense cDNA of MMP-9 was constructed and transfected into a non-metastatic human melanoma cell line WM35, which does not express MMP-9. In vitro test such as growth rate, MTT method, 3H-thymidine incorporation, colony forming of soft agar,Boyden chamber, as well as Zymography and Western Blot experiment were used to analyse expression of MMPs and in vitro behavior of tumor cells before and after gene transfection.Results After transfection of sense MMP-9 the expression and activity of MMP-9 increased, the capacity of in vitro growth,invasion through Matrigel was raised to some extent.Conclusion Transfection of sense MMP-9 enhanced capacity of growth and invasion.