USE OF IMMUNOLOGICAL TECHNIQUES TO STUDY THE INTERACTION OF CARCINOGENS WITH DNA

针对致癌物-DNA加合物的抗体已经探索了这种性质， 体外和体内DNA修饰的程度和后果。DNA 取代为2-乙酰氨基荧烯(AAF)和 对顺二亚胺氯铂II(cis-DDP)进行了定量分析 每个细胞能够检测约100个加合物的免疫分析方法 发展了免疫组织化学程序来定位加合物的原位。在……里面 给予致癌剂量AAF 4周大鼠肝DNA加合物 积累量在2-3周时达到高峰。因为加合物去除是 在随后的4周内进行双相控制饮食，由计算机得出 药代动力学模型认为加合物形成两个基因组 一个隔间，它们被迅速从一个隔间移走，另一个隔间 它们被慢慢地移走。为识别这两个人而启动的研究 间隔室涉及不同类型肝细胞的加合物分析和 不同的染色质区域，包括核基质。 饲喂大鼠肝脏内AF-DNA加合物的免疫组织化学定位 荧光和显微荧光法显示AAF是浓缩的。 主要发生在门脉周围区域。此外，没有加成物。 免疫荧光技术在几种致癌前病变中的检测 不同的协议。有核的外周血细胞DNA从 癌症患者在顺式顺铂治疗过程中的多次，以及 共对254个样品进行了分析。其中，23个未经处理的对照样本 在231例接受顺铂治疗的患者中，43%的样本为阴性 都是阳性。在阳性患者中，加合物积聚以一种 总累积剂量的函数，暗示相对较慢的加合 移走。55名患者的疾病反应数据表明，个人 加合物水平高的患者对治疗的完全应答率很高。 在动物模型中的平行实验已经证明了同样的 在大鼠和小鼠的肾脏、性腺和肿瘤中直接形成加合物形式 与剂量的关系。

Antibodies specific for carcinogen-DNA adducts have probed the nature, extent, and consequences of in vitro and in vivo DNA modification. DNAs substituted with 2-acetylaminofluorene (AAF) and cis-diamiminedichloroplatinum II (cis-DDP) were analyzed by quantitative immunoassays able to detect about 100 adducts per cell and by immunohistochemical procedures developed to localize adducts in situ. In hepatic DNA of rats fed a carcinogenic dose of AAF for 4 weeks, adduct accumulation reached a plateau at 2-3 weeks. Since adduct removal was biphasic during 4 subsequent weeks on control diet, a computer-derived pharmacokinetic model proposed that adducts are formed into two genomic compartments, one from which they are removed rapidly and another from which they are removed slowly. Studies initiated to identify these two compartments involve adduct analysis in different liver cell types and different chromatin regions including the nuclear matrix. Immunohistochemical localization of AF-DNA adducts in livers of rats fed AAF was shown by fluorescence and microfluorometry to be concentrated primarily in periportal regions. In addition, there were no adducts detectable by immunofluorescence in preneoplastic foci induced by several different protocols. Nucleated peripheral blood cell DNA was obtained from cancer patients at multiple times during courses of cis-DDP therapy, and a total of 254 samples were analyzed. Of these, 23 untreated control samples were negative, and 43% of the 231 samples from patients receiving cis-DDP were positive. Adduct accumulation, in positive patients, occurred as a function of total cumulative dose, suggesting relatively slow adduct removal. Disease response data on 55 patients indicated that individuals with high adduct levels have a high rate of complete response to therapy. Parallel experiments in animal models have demonstrated that the same adduct forms in kidney, gonads, and tumors of rats and mice in direct relation to dose.