Champion A R, Hanson J A, Court J B, Venables S E
Research Laboratories, Velindre NHS Trust, Whitchurch, Cardiff, UK.
Mutagenesis. 1995 May;10(3):203-8. doi: 10.1093/mutage/10.3.203.
The cytokinesis-block micronucleus assay was used to measure radiosensitivity in vitro in a panel of seven cell lines. Six of these cell lines were used to study the major parameters of this assay. We observed varying sensitivities following cytochalasin-B exposure. Treatment with 1 microgram/ml cytochalasin-B for 24 h reduced cell survival in four of the six cell lines by > 60%. Cytochalasin-B concentration and post-irradiation culture time were both found to influence cell-response. In three cell lines (V39, V134 and HX142), a decrease in cytochalasin-B concentration (2-0.5 microgram/ml) resulted in an increase in the frequency of radiation-induced micronuclei per binucleate cell. In other cell lines, either the opposite (V7M, CHO-K1) or no effect (WiDr) was seen. A linear dose-response was observed between induced damage expressed as the frequency of micronuclei and radiation dose in all but one melanoma (V39) cell line. Evidence for radiation-induced division-delay, with the maximum frequency of binucleation in irradiated cultures occurring 24-48 h after that of controls, was only seen in two cell lines. Of particular note, and in contrast to some other published reports, was the lack of a general correlation between cell-response measured in the clonogenic and the cytokinesis-block micronucleus assays. Consideration of lethal lesions, determined from the clonogenic dose-response curve, with respect to micronucleus frequency showed a complex relationship, with one micronucleus per binucleate cell corresponding to a wide range of lethal lesions depending on the cell line.(ABSTRACT TRUNCATED AT 250 WORDS)
胞质分裂阻断微核试验用于在一组七种细胞系中体外测量放射敏感性。其中六种细胞系用于研究该试验的主要参数。我们观察到细胞松弛素-B处理后敏感性存在差异。用1微克/毫升细胞松弛素-B处理24小时,六种细胞系中的四种细胞系的细胞存活率降低超过60%。发现细胞松弛素-B浓度和照射后培养时间均会影响细胞反应。在三种细胞系(V39、V134和HX142)中,细胞松弛素-B浓度降低(从2微克/毫升降至0.5微克/毫升)导致每个双核细胞中辐射诱导微核的频率增加。在其他细胞系中,观察到相反的情况(V7M、CHO-K1)或无影响(WiDr)。除一种黑色素瘤(V39)细胞系外,在所有细胞系中,以微核频率表示的诱导损伤与辐射剂量之间均观察到线性剂量反应。仅在两种细胞系中发现辐射诱导分裂延迟的证据,即照射培养物中双核化的最大频率出现在对照之后的24 - 48小时。特别值得注意的是,与其他一些已发表的报告相反,克隆形成试验和胞质分裂阻断微核试验所测量的细胞反应之间缺乏普遍相关性。根据克隆形成剂量反应曲线确定致死损伤,并考虑微核频率,结果显示存在复杂关系,每个双核细胞中有一个微核对应于取决于细胞系的广泛致死损伤范围。(摘要截短于250字)