Detection of hypoxia by measurement of DNA damage in individual cells from spheroids and murine tumours exposed to bioreductive drugs. II. RSU 1069.
Detection of hypoxia by measurement of DNA damage in individual cells from spheroids and murine tumours exposed to bioreductive drugs. II. RSU 1069.
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DOI：
10.1038/bjc.1995.106
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发表时间：
1995-03
影响因子：
8.8
通讯作者：
Olive, P L
中科院分区：
文献类型：
--
作者：
Olive, P L
双功能生物还原药物RSU 1069识别多细胞球体和小鼠SCCVII鳞状细胞癌中缺氧细胞的能力使用碱性彗星法进行了检查。该方法应用荧光显微镜和图像分析来测量包埋在琼脂糖中并暴露于电场的单个细胞的DNA迁移量。将暴露于RSU 1069 1 h的中国仓鼠V79球状体解聚，并分析单个细胞的DNA损伤。暴露于RSU 1069后，需氧细胞表现出DNA单链断裂，而缺氧细胞中产生DNA链间交联。含有40-50%放射生物学低氧细胞的球状体表现出20-30%的细胞具有交联，其余的仅显示出链断裂。在暴露于25-200 mg kg-1的C3 H小鼠中生长的SCCVII肿瘤中观察到类似的损伤模式。随后在体外照射细胞，大大提高了区分从球状体或SCCVII小鼠肿瘤暴露于RSU 1069，特别是在低剂量的药物治疗后，好氧和缺氧细胞。损伤模式在药物注射后至少4 h相对稳定。结果表明，检测实体瘤中的缺氧细胞可能是实用的，使用这种代理或前药，PD 144872，选择用于I期临床试验作为人类肿瘤中的缺氧细胞放射增敏剂和细胞毒素。
The ability of the dual-function bioreductive drug, RSU 1069, to identify hypoxic cells in multicell spheroids and murine SCCVII squamous cell carcinomas was examined using the alkaline comet method. This method applies fluorescence microscopy and image analysis to measure the amount of migration of DNA from individual cells embedded in agarose and exposed to an electric field. Chinese hamster V79 spheroids, exposed for 1 h to RSU 1069, were disaggregated and individual cells were analysed for DNA damage. Following exposure to RSU 1069, aerobic cells exhibited DNA single-strand breaks while DNA interstrand cross-links were produced in hypoxic cells. Spheroids containing 40-50% radiobiologically hypoxic cells exhibited 20-30% cells with cross-links and the remainder showed only strand breaks. Similar patterns of damage were observed in SCCVII tumours growing in C3H mice exposed to 25-200 mg kg-1. Subsequent irradiation of cells in vitro greatly improved the distinction between aerobic and hypoxic cells from spheroids or SCCVII murine tumours exposed to RSU 1069, especially after treatment with low drug doses. The pattern of damage was relatively stable for at least 4 h after drug injection. Results indicate that detection of hypoxic cells in solid tumours may be practical using this agent or a prodrug, PD 144872, selected for phase I clinical testing as a hypoxic cell radiosensitiser and cytotoxin in human tumours.