TRANSFECTION OF C6 GLIOMA-CELLS WITH CONNEXIN-43 CDNA - ANALYSIS OF EXPRESSION, INTERCELLULAR COUPLING, AND CELL-PROLIFERATION
TRANSFECTION OF C6 GLIOMA-CELLS WITH CONNEXIN-43 CDNA - ANALYSIS OF EXPRESSION, INTERCELLULAR COUPLING, AND CELL-PROLIFERATION
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DOI：
10.1073/pnas.88.5.1883
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发表时间：
1991-03-01
影响因子：
11.1
通讯作者：
NAUS, CCG
中科院分区：
文献类型：
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作者：
ZHU, D;CAVENEY, S;NAUS, CCG
C6胶质瘤细胞表达低水平的差距连接蛋白连接蛋白43及其mRNA，并显示非常弱的染料偶联。 当植入大鼠大脑时，这些细胞迅速产生一个大的神经胶质瘤。 为了研究间隙连接在这些细胞的肿瘤特征中的作用，我们使用脂质体介导的转染引入编码连接蛋白43的全长cDNA。 获得了几个转染的克隆，其表现出从插入的cDNA转录的不同量的连接蛋白43 mRNA。 免疫细胞化学分析显示，在转染细胞中连接蛋白43免疫反应性的量增加，被定位在细胞间接触的区域，以及在细胞质中。 还评估了染料偶联的水平，发现其与连接蛋白43 mRNA的量相关。 当细胞增殖几天后，表达转染的cDNA的细胞比未转染的细胞生长得更慢。 这些转染的细胞将有助于研究间隙连接在肿瘤发生中的作用。
C6 glioma cells express low levels of the gap junction protein connexin 43 and its mRNA and display very weak dye coupling. When implanted into the rat cerebrum, these cells quickly give rise to a large glioma. To investigate the role of gap junctions in the tumor characteristics of these cells, we have used Lipofectin-mediated transfection to introduce a full-length cDNA encoding connexin 43. Several transfected clones were obtained that exhibited various amounts of connexin 43 mRNA transcribed from the inserted cDNA. Immunocytochemical analysis revealed an increase in the amount of connexin 43 immunoreactivity in the transfected cells, being localized at areas of intercellular contact as well as in the cytoplasm. The level of dye coupling was also assessed and found to correlate with the amount of connexin 43 mRNA. When cell proliferation was followed over several days, cells expressing the transfected cDNA grew more slowly than nontransfected cells. These transfected cells will be useful in examining the role of gap junctions in tumorigenesis.