Immunolocalization of 8-Oxoguanine in Nutrient-Deprived Mammalian Tissue
Immunolocalization of 8-Oxoguanine in Nutrient-Deprived Mammalian Tissue
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营养缺乏的哺乳动物组织中 8-氧代鸟嘌呤的免疫定位
DOI：
10.1179/his.2000.23.1.37
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发表时间：
2000
影响因子：
1.1
通讯作者：
M. Berrios
中科院分区：
文献类型：
--
作者：
Kimberly A. Conion;A. Grollman;M. Berrios
摘要组织在缺血和其他形式的损伤中所经历的营养剥夺可能会导致DNA氧化损伤。为了探索这种可能性，对缺乏营养的人HeLa和猿猴Cos7细胞进行了8-氧鸟嘌呤的评估，8-氧鸟嘌呤是一种公认的DNA氧化损伤的标志。用抗8-氧鸟嘌呤的单抗在体外和原位检测和定位这一病变。在受控培养条件下。短期的营养剥夺足以诱导活跃生长的HeLa和Cos7细胞中8-氧鸟嘌呤的积累。免疫荧光分析表明，在营养缺乏的细胞中，8-氧鸟嘌呤主要定位于细胞核，一旦细胞恢复正常生长条件，这种核聚集可以逆转到基础水平。此外，抗8-羟鸟嘌呤抗体的共聚焦免疫荧光显示，在营养缺乏的细胞中，间期核的外围和内部有几个明显的强染色区域。使用这一新的方法，我们发现相对温和的内源性和外源性因素对DNA氧化损伤有贡献。因为8-氧鸟嘌呤具有致突变性，我们相信进一步的证据可能提供证据支持内源性DNA损伤与退行性疾病和癌症的病因有关的假说。(《组织技术评论》23：37,2000)
Abstract Nutrient deprivation, as experienced by tissues during ischemia and other forms of injury may induce oxidative DNA damage. To explore this possibility, nutrient-deprived human HeLa and simian Cos7 cells were evaluated for 8-oxoguanine, a well established marker for oxidative DNA damage. Monoclonal antibodies directed against 8-oxoguanine were used to detect and localize this lesion in vitro and in situ. Under controlled culture conditions. short term nutrient deprivation was sufficient to induce the accumulation of 8-oxoguanine in actively growing HeLa and Cos7 cells. Immunofluorescencaen alyses revealed that 8-oxoguanine was primarily localized to the nucleus in nutrientdeprived cells, and this nuclear accumulation could be reversed to base levels once cells were returned to normal growing conditions. Furthermore,confocal immunofluorescence with anti-8-oxoguanine antibodies revealed several distinct areas of intense staining at the periphery and interior of interphase nuclei in nutrient deprived cells. Using this novel approach, we have found that relatively mild endogenous and exogenous factors contribute to oxidative DNA damage. Because 8-oxoguanine is mutagenic, we believe furthers tudies may provide evidence to support the hypothesis that endogenous DNA damage is involved in the etiology of both degenerative diseases and cancer. (The J Histotechnol 23:37, 2000)
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DOI：
10.1002/(sici)1520-6327(1998)37:3
发表时间：
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期刊：
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影响因子：
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