Reduction of drug accumulation and DNA topoisomerase II activity in acquired teniposide-resistant human cancer KB cell lines.
Reduction of drug accumulation and DNA topoisomerase II activity in acquired teniposide-resistant human cancer KB cell lines.
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获得性替尼泊苷耐药人类癌症 KB 细胞系中药物蓄积和 DNA 拓扑异构酶 II 活性的降低。
作者：
Ken;K. Kohno;H. Takano;Shin;A. Kiue;M. Kuwano
我们通过逐步增加药物剂量，从人类癌症 KB 细胞中分离出稳定的替尼泊苷 (VM26) 耐药细胞系。在每一步中，我们都纯化了 VM26 抗性细胞系。 KB/VM-a、KB/VM-b、KB/VM-1、KB/VM-2、KB/VM-3 和 KB/VM-4 对 VM26 的耐药性比 KB 高 3、6、12、16、74 和 95 倍。我们进一步表征了 KB/VM-2 和 KB/VM-4，它们对 VM26 或依托泊苷 (VP16) 的耐药性比 KB 高约 15 倍和 100 倍。两种 VM26 耐药细胞系对道诺霉素和阿霉素的相对耐药性比 KB 高 4 至 11 倍。 KB/VM-2和KB/VM-4细胞中放射性VP16细胞累积的稳态水平约为KB细胞的40%，而KB/VM-2和KB/VM-4细胞中观察到的放射性道诺霉素累积水平与KB细胞相似。通过动质体 DNA 的串联测定，KB/VM-2 和 KB/VM-4 的核提取物的拓扑异构酶 II 活性始终是 KB 细胞活性的三分之二或更少。在使用特异性抗拓扑异构酶 II 抗体的免疫印迹分析和使用特定人 DNA 拓扑异构酶 II 互补 DNA 的 Northern 印迹分析中都观察到类似的减少。然而，突变体与其亲本之间的 DNA 拓扑异构酶 I 活性相似。此外，KB/VM-2和KB/VM-4的细胞生长比KB更不耐热，而KB/VM-b已经表现出对温度敏感的生长。与 KB/VM-2 或 KB/VM-4 一样，KB/VM-1 确实显示出 VP16 积累减少，但与 KB 细胞一样，其拓扑异构酶 II 含量处于正常水平。这些数据表明，DNA 拓扑异构酶 II 表达的减少，可能与药物通透性降低相结合，可以解释 KB/VM-2 和 KB/VM-4 细胞获得性 VM26 耐药性，而且温度敏感表型可能不一定与拓扑异构酶 II 表达减少或通透性降低相关。
We have isolated stable teniposide (VM26)-resistant cell lines from human cancer KB cells by stepwise exposure to increasing doses of the drug. At each step, we have purified VM26-resistant cell lines. KB/VM-a, KB/VM-b, KB/VM-1, KB/VM-2, KB/VM-3, and KB/VM-4 showed 3-, 6-, 12-, 16-, 74-, and 95-fold higher resistance to VM26 than did KB. We have further characterized KB/VM-2 and KB/VM-4 which showed about 15- and 100-fold higher resistance to VM26 or etoposide (VP16) than did KB. Both VM26-resistant cell lines showed 4- to 11-fold higher relative resistance to daunomycin and Adriamycin than did KB. Steady-state levels of the cellular accumulation of radioactive VP16 in KB/VM-2 and KB/VM-4 cells were about 40% of that of KB cells, whereas similar levels of radioactive daunomycin accumulation were observed in KB/VM-2 and KB/VM-4 cells as KB cells. Topoisomerase II activity of nuclear extracts of both KB/VM-2 and KB/VM-4 assayed by decatenation of kinetoplast DNA was consistently two-thirds or less the activity of KB cells. A similar reduction was seen in both immunoblot assays with specific anti-topoisomerase II antibody and Northern blot analysis with specific human DNA topoisomerase II complementary DNA. DNA topoisomerase I activity, however, was similar between the mutants and their parent. Furthermore, cell growth of KB/VM-2 and KB/VM-4 was more thermolabile than that of KB, while KB/VM-b already showed temperature-sensitive growth. KB/VM-1 did show reduced accumulation of VP16 as in KB/VM-2 or KB/VM-4, but it had a normal level of topoisomerase II content as in KB cells. These data suggest that the reduced expression of DNA topoisomerase II, possibly combined with decreased permeability to the drugs, can account for the acquired VM26 resistance of KB/VM-2 and KB/VM-4 cells and also that the temperature-sensitive phenotype might not be obligatorily coupled with the reduced expression of topoisomerase II or the decreased permeability.
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影响因子：
11.2
作者：
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来自 HeLa 细胞核的同质 II 型 DNA 拓扑异构酶。
DOI：
--
发表时间：
1981
期刊：
The Journal of biological chemistry
影响因子：
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DOI：
10.1016/s0021-9258(17)47282-6
发表时间：
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期刊：
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影响因子：
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