【摘要】 目的 :探讨金属蛋白酶MMP 9与肿瘤恶性表型的相关性以及其可调控型表达在肿瘤基因治疗中的应用。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 :在外源性四环素存在时 ,转染基因不表达 ,细胞表型无明显变化。去除四环素 ,WM35细胞MMP 9的表达及活性增强 ,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。结论 :正义MMP 9基因的可调控型表达能促进人黑色素瘤细胞的生长和侵袭 ,进一步证明MMP 9在人黑色素瘤细胞侵袭过程中起重要作用。更多还原

【Abstract】 Objective: To investigate the correlation between matrix metalloproteinase 9 (MMP 9) expression and tumor metastasis, and explore the potential application of controlled expression of target gene in tumor gene therapy. Methods:One self contained tetracycline regulated retroviral vector containing sense cDNA of MMP 9 was constructed and transfected into an early stage human melanoma cell line WM35, which did not express MMP 9. In vitro tests such as growth rate, MTT method, 3 H thymidine incorporation, colony forming ability in soft agar, in vitro invasion assay in Boyden chambers, as well as zymography and Western Blot experiment were used to analyze expression of MMPs and in vitro behavior of tumor cells before and after gene transfection. Results: In the presence of exogenous tetracycline, the expression of the transfected MMP 9 were under detectable level and no significant changes in cell behaviors were found when compared with vector transfected control cells. But when the tetracycline was withdrew from the medium, the expression and activity of MMP 9 were significantly increased. The capacity of in vitro growth, colony forming ability in soft agar, invasion through Matrigel were enhanced remarkably. Conclusion: Transfection of sense MMP 9 can enhance growth and invasion of melanoma cells, further confirming its important role in tumor invasion and metastasis.更多还原