Terheggen P M, Emondt J Y, Floot B G, Dijkman R, Schrier P I, den Engelse L, Begg A C
Division of Chemical Carcinogenesis, The Netherlands Cancer Institute, (Antoni van Leeuwenhoek Huis), Amsterdam.
Cancer Res. 1990 Jun 15;50(12):3556-61.
The relationship between cell killing and the binding of the anticancer drug cis-diamminedichloroplatinum(II) (cis-DDP) to DNA was studied in six mammalian cell lines. Two of the human cell lines (COV413B) were of the same origin, comprising one sensitive to cis-DDP and the other with induced resistance to the drug. The four other lines, two rodent (RIF-1, Chinese hamster ovary) and two human (A2780, A1847), were unrelated. The cell lines differed in their sensitivity to cis-DDP, as tested in a clonogenic assay. cis-DDP-DNA binding was determined by quantitative immunocytochemistry using an antiserum against cis-DDP-modified DNA. The resistance factors relative to RIF-1, calculated from full survival curves for cis-DDP, were 3.8 +/- 0.4 for Chinese hamster ovary cells and 8.8 +/- 0.7 for both A2780 and A1847 lines. Using quantitative immunocytochemistry, the levels of the adduct-specific nuclear staining density compared with RIF-1 cells were 4.8 +/- 0.2 for Chinese hamster ovary cells, 9.1 +/- 0.2 for A2780, and 10.0 +/- 0.1 for A1847 cells, i.e., in good agreement with the resistance factors. In studies with the COV413B cells and their cis-DDP-resistant counterpart COV413B-PtR, immunologically detected adduct levels again correlated closely with resistance factors (correlation coefficient = 0.97). The kinetics of cis-DDP-DNA adduct formation and loss was investigated in RIF-1, A2780, and A1847 cells by the immunocytochemistry technique. Adduct levels after a 1-h incubation with approximately equitoxic doses of cis-DDP increased by 18 to 32% (average, 27%) between 0 and 6.5 h after treatment and then declined. Adduct half-lives in this latter phase did not correlate with the sensitivities of the cells for cis-DDP. These results indicate that the initial level of cis-DDP-DNA binding measured by quantitative immunocytochemistry may be a reasonable predictor of sensitivity to this chemotherapeutic drug.
在六种哺乳动物细胞系中研究了细胞杀伤与抗癌药物顺二氨二氯铂（II）（顺铂）与DNA结合之间的关系。两种人类细胞系（COV413B）来源相同，其中一种对顺铂敏感，另一种对该药物具有诱导抗性。另外四种细胞系，两种啮齿动物（RIF-1、中国仓鼠卵巢细胞）和两种人类（A2780、A1847），彼此无关。通过克隆形成试验测试，这些细胞系对顺铂的敏感性不同。使用抗顺铂修饰DNA的抗血清，通过定量免疫细胞化学测定顺铂与DNA的结合。根据顺铂的完整存活曲线计算，相对于RIF-1的抗性因子，中国仓鼠卵巢细胞为3.8±0.4，A2780和A1847细胞系均为8.8±0.7。使用定量免疫细胞化学，与RIF-1细胞相比，中国仓鼠卵巢细胞加合物特异性核染色密度水平为4.8±0.2，A2780为9.1±0.2，A1847细胞为10.0±0.1，即与抗性因子高度一致。在对COV413B细胞及其顺铂抗性对应物COV413B-PtR的研究中，免疫检测到的加合物水平再次与抗性因子密切相关（相关系数=0.97）。通过免疫细胞化学技术研究了RIF-1、A2780和A1847细胞中顺铂-DNA加合物形成和消失的动力学。用大约等毒性剂量的顺铂孵育1小时后，加合物水平在处理后0至6.5小时之间增加了18%至32%（平均27%），然后下降。后一阶段加合物的半衰期与细胞对顺铂的敏感性无关。这些结果表明，通过定量免疫细胞化学测量的顺铂-DNA结合初始水平可能是对这种化疗药物敏感性的合理预测指标。
