Selective suppression of matrix metalloproteinase-9 in human glioblastoma cells by antisense gene transfer impairs glioblastoma cell invasion.
Selective suppression of matrix metalloproteinase-9 in human glioblastoma cells by antisense gene transfer impairs glioblastoma cell invasion.
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发表时间：
2000-12
期刊：
影响因子：
11.2
通讯作者：
S. Kondraganti;S. Mohanam;S. Chintala;Y. Kin;S. Jasti;C. Nirmala;S. Lakka;Y. Adachi;A. Kyritsis-A.-K
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文献类型：
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作者：
S. Kondraganti;S. Mohanam;S. Chintala;Y. Kin;S. Jasti;C. Nirmala;S. Lakka;Y. Adachi;A. Kyritsis-A.-K
基质金属蛋白酶(MMPs)的高表达与人脑胶质母细胞瘤的肿瘤进展有关。在这项研究中，我们试图通过稳定地将能够表达与人基质金属蛋白酶-9基因5‘端528-bp片段互补的反义转录的质粒载体导入高级别胶质母细胞瘤细胞系来下调基质金属蛋白酶-9的表达。经G418筛选，获得稳定的转染体。在载体、正义和反义载体的克隆中，Northern印迹、Western blotting和明胶酶谱显示，只有反义转染的细胞中，基质金属蛋白酶-9的表达显著降低。Matrigel侵袭实验显示，与亲本克隆、载体克隆和正义克隆相比，反义克隆的侵袭力显著降低。肿瘤球体和胎鼠脑集合体的共培养表明，反义转染的稳定克隆没有显示出对大鼠脑集合体的侵袭；相比之下，90%的亲本克隆、载体克隆和正义克隆侵袭了大鼠脑集合体。在裸鼠脑内注射反义稳定基因没有产生肿瘤或非常小的肿瘤，但在脑内注射亲本或载体克隆确实产生了肿瘤。这些结果提示，基质金属蛋白酶-9的表达在胶质母细胞瘤细胞的侵袭性中起重要作用。
Increased expression of matrix metalloproteinases (MMPs) has been associated with human glioblastoma tumor progression. In this study, we sought to down-regulate MMP-9 expression by stably transfecting a high-grade glioblastoma cell line with a plasmid vector capable of expressing an antisense transcript complementary to a 528-bp segment at the 5' end of human MMP-9 cDNA. Stable transfectants were obtained through selection with G418. Of the clones transfected with vector, sense, and antisense constructs, Northern blotting, Western blotting, and gelatin zymography showed that MMP-9 expression was significantly reduced only in the antisense-transfected cells. A Matrigel invasion assay revealed marked reductions in invasiveness for the antisense clones relative to the parental, vector, and sense clones. Cocultures of tumor spheroids and fetal rat brain aggregates showed that the antisense-transfected stable clones showed no invasion of the rat brain aggregates; in contrast, 90% of the parental, vector, and sense clones invaded the rat brain aggregates. Intracerebral injection of antisense stable transfectants in nude mice produced no tumors or very small tumors, but intracerebral injection of parental or vector clones did produce tumors. These results suggest that MMP-9 expression is essential for the invasiveness of glioblastoma cells.