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目的探讨金属蛋白酶MMP-9与肿瘤转移的相关性。方法利用基因重组技术构建反义MMP-9cDNA四环素可调控表达载体,用脂质体法转染反义MMP-9至转移性人黑色素瘤细胞株WM451(高表达MMP-9)。检测转染后细胞MMP-9表达水平的改变以及裸鼠体内成瘤及自发转移能力的变化。结果转染反义基因后,WM451细胞MMP-9的表达及活性明显下降,体外侵袭能力及裸鼠体内成瘤性及自发转移能力均受到一定程度抑制;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。结论反义MMP-9基因下调MMP-9的表达,可使人黑色素细胞转移能力受到一定程度的抑制,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。
Objective To investigate the correlation between metalloproteinase MMP-9 and tumor metastasis. Methods Antisense MMP-9 cDNA tetracycline-regulated expression vector was constructed by gene recombination technique. Antisense MMP-9 was transfected into human melanoma cell line WM451 (overexpression MMP-9) by lipofectamine. The changes of MMP-9 expression in transfected cells and the ability of tumorigenesis and spontaneous metastasis in nude mice were detected. Results After transfected with antisense gene, the expression and activity of MMP-9 in WM451 cells were significantly decreased. The invasion ability in vitro and the ability of tumorigenesis and spontaneous metastasis in vivo were inhibited to a certain extent. Tetracycline inhibited the expression of tetracycline-negative retrovirus Expression of exogenous genes on vectors. Conclusion The down-regulation of MMP-9 expression by antisense MMP-9 gene can inhibit the metastasis of human melanocytes to a certain extent, indicating that MMP-9 plays an important role in the metastasis of human melanoma cells. Meanwhile, tetracycline negative regulatory retroviral vectors can regulate the expression of foreign genes.