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目的 探讨基质金属蛋白酶 (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四环素可调控型表达载体 ,用脂质体法转染反义MMP 9至转移性人黑色素瘤细胞株WM4 5 1(高表达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、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 ,WM4 5 1细胞MMP 9的表达及活性明显下降 ,同时MMP 2的表达也受到一定抑制 ,细胞生长速度、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 relationship between matrix metalloproteinase (MMP) 9 and tumor metastasis. Methods Antisense MMP 9 cDNA tetracycline regulatable expression vector was constructed by gene recombination technique. Antisense MMP 9 was transfected into human melanoma cell line WM4 5 1 (high expression of MMP 9) by liposome method. The expression of MMP-9 in transfected cells was detected and the growth, invasion, tumor formation and spontaneous metastasis in nude mice were observed. Results After transfection of antisense gene, the expression and activity of MMP-9 in WM4 5 1 cells were significantly decreased. Meanwhile, the expression of MMP-2 was also inhibited. The growth rate, invasiveness in vitro, tumorigenicity and spontaneous metastasis in nude mice Was inhibited to a certain extent; the use of tetracycline could inhibit the expression of foreign genes on the tetracycline negative regulation retroviral vector. Conclusion The downregulation 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 vector can regulate the expression of foreign genes