Olive P L, Banáth J P, MacPhail H S
British Columbia Cancer Research Centre, Vancouver, Canada.
Cancer Res. 1994 Jul 15;54(14):3939-46.
The neutral filter elution method and the neutral comet assay have been used to analyze radiation-induced DNA damage and repair in 6 human tumor cell lines: HT-144 melanoma; DU-145 prostate carcinoma; U-87 glioma; WiDr and HT-29 colon adenocarcinomas; and SiHa cervical carcinoma. In spite of large differences in intrinsic radiosensitivity measured using a clonogenic assay, double-strand break induction, rejoining rate, and amount of residual DNA damage 4 h after irradiation were similar among these cell lines when measured using the neutral comet assay. Differences in initial numbers of double-strand breaks were observed using the neutral filter elution method; however, there was no correlation with radiosensitivity, nor did the rejoining rate or amount of residual DNA damage measured using filter elution correlate with radiation sensitivity. We conclude that neither double-strand break assay is able to reliably rank cells according to clonogenic survival following irradiation.
中性滤膜洗脱法和中性彗星试验已用于分析6种人类肿瘤细胞系中辐射诱导的DNA损伤和修复情况:HT-144黑色素瘤细胞系;DU-145前列腺癌细胞系;U-87胶质瘤细胞系;WiDr和HT-29结肠腺癌细胞系;以及SiHa宫颈癌细胞系。尽管使用克隆形成试验测得的内在放射敏感性存在很大差异,但当使用中性彗星试验测量时,这些细胞系在照射后4小时的双链断裂诱导、重新连接率和残留DNA损伤量相似。使用中性滤膜洗脱法观察到双链断裂初始数量存在差异;然而,这与放射敏感性无关,并且使用滤膜洗脱法测量的重新连接率或残留DNA损伤量也与辐射敏感性无关。我们得出结论,两种双链断裂试验均无法根据照射后的克隆形成存活率可靠地对细胞进行排名。