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Induced Synthesis of O6‐Methylguanine‐DNA Methyltransferase in Rat Hepatoma Cells Exposed to DNA‐damaging Agents

机译:暴露于DNA破坏剂的大鼠肝细胞中O6-甲基鸟嘌呤-DNA甲基转移酶的诱导合成

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摘要

When the rat hepatoma cell line H4IIE was treated with DNA‐damaging agents such as N‐methyl‐N′‐nitro‐N‐nitrosoguanidine (MNNG), ultraviolet light and γ‐rays, the O6‐methylguanine‐DNA methyltransferase activity increased 2 to 3 times over the level seen in non‐treated cells. SDS/polyacrylamide gel electrophoresis followed by fluorography revealed that a single species of methyltransferase protein with a molecular weight of 25,500 was present in both non‐treated and treated cells. Northern blot analysis using a cloned rat cDNA as a probe revealed that the enzyme activity increased because transcription of the gene was enhanced. The level of enzyme activity increased within 48 h after UV irradiation and remained at a higher level for 150 h. Following UV irradiation, the cells become more resistant than the normal cells to MNNG.
机译:当用DNA破坏剂如N-甲基-N'-硝基-N-亚硝基胍(MNNG),紫外线和γ射线处理大鼠肝癌细胞H4IIE时,O 6 -甲基鸟嘌呤-DNA甲基转移酶活性比未处理的细胞提高了2到3倍。 SDS /聚丙烯酰胺凝胶电泳,然后进行荧光分析,发现未处理和处理过的细胞中均存在一种分子量为25,500的甲基转移酶蛋白。使用克隆的大鼠cDNA作为探针的Northern印迹分析表明,酶活性增加是因为该基因的转录增强了。酶活性水平在紫外线照射后48小时内增加,并在150小时内保持较高水平。紫外线照射后,细胞比正常细胞对MNNG的抵抗力更高。

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