Pintzas A, Spandidos D A
Hellenic Pasteur Institute, Athens, Greece.
Gene Anal Tech. 1989 Nov-Dec;6(6):125-30. doi: 10.1016/0735-0651(89)90003-4.
Rat fibroblast cells carrying an exogenous normal or mutant T24 human H-ras1 gene were transfected with plasmids carrying the normal or mutant T24 H-ras1 gene promoter linked to the reporter chloramphenicol acetyl transferase (CAT) gene and the cells were treated with insulin. We found that the H-ras1 gene was positively autoregulated and that insulin potentiated the response of the T24 ras p21 to the H-ras1 gene promoter. We have also examined the effect of insulin directly on the H-ras1 promoter by treating stable transfectants obtained after transfection of rat fibroblasts with plasmids carrying the normal or mutant T24 H-ras1 gene promoter linked to the reporter CAT gene and the selectable marker aminoglycoside phosphotransferase (aph) gene. We found that insulin appeared to have no direct effect on the H-ras1 promoter in this case, suggesting that the effect is mediated through the ras p21 oncogene product. We suggest that the mutant T24 H-ras p21 protein mediates the action of insulin.
将携带外源性正常或突变型T24人H-ras1基因的大鼠成纤维细胞用携带与报告氯霉素乙酰转移酶（CAT）基因相连的正常或突变型T24 H-ras1基因启动子的质粒进行转染，然后用胰岛素处理这些细胞。我们发现H-ras1基因存在正性自我调节，并且胰岛素增强了T24 ras p21对H-ras1基因启动子的反应。我们还通过用携带与报告CAT基因和选择标记氨基糖苷磷酸转移酶（aph）基因相连的正常或突变型T24 H-ras1基因启动子的质粒转染大鼠成纤维细胞后获得的稳定转染子，直接检测了胰岛素对H-ras1启动子的影响。我们发现在这种情况下胰岛素似乎对H-ras1启动子没有直接影响，这表明该效应是通过ras p21癌基因产物介导的。我们认为突变型T24 H-ras p21蛋白介导了胰岛素的作用。
