FLOW CYTOMETRIC STUDY OF ANTICANCER DRUG EFFECTS ON DNA

这项研究的总体目标是开发一种预测性的 一种可选择有效的肿瘤烷化剂的检测方法 治疗和检测耐药肿瘤细胞。最新进展 的化验结果将基于o之间的线性关系 单抗F7-26与细胞DNA的结合 氮芥衍生物和亚硝脲对小鼠的细胞毒性 1-6对数细胞杀伤率范围；单抗的杀伤能力 区分敏感细胞和耐药细胞。少校 在这项研究中需要回答的问题是，量化的 单抗结合与药物敏感性或耐药性的相关性 可以在不同的小鼠和人类肿瘤细胞中观察到。单抗 单个细胞中与DNA的结合将通过Flow来测量 细胞学。单抗F7-26免疫反应性与乳腺癌的相关性 烷化剂处理细胞中的DNA与细胞杀伤 将在两对敏感和耐药的细胞系中进行研究 对烷化剂的作用：1)肺癌A549(Mer+ 表型)和A427(Mer表型)对 2)卵巢癌细胞株A2780 对L-PAM敏感和耐药。在每一对线路中， 细胞存活、DNA初始损伤与修复的关系 将对药物移除后的DNA损伤进行研究。鼠标实体 肿瘤Lewis肺癌及其耐药亚系 亚硝脲和环磷酰胺将在#年用药治疗 体外和体内。单抗与异倍体肿瘤细胞的结合 将其与肿瘤治疗反应进行比较。这个 低DNA细胞亚群的出现与其关系的研究 免疫反应性与耐药的发生发展 重复的药物治疗将被研究。来描述类型的特征 用抗原决定簇单抗检测DNA的变化 F7-26单抗的研究将采用直接和竞争性的酶联免疫吸附试验。 免疫分析。单抗对不同细胞株的相对亲和力 脱氧核均聚物与寡核苷酸中的碱基数目 结合单抗所需的条件将会确定。流式细胞仪 建议研究药物对单个肿瘤细胞DNA的影响 在这里应该可以将新的方法应用于 临床环境。

The general goal of this study is the development of a predictive assay which may select effective alkylating agents for tumor treatment and detect drug-resistant tumor cells. The development of the assay will be based on o linear relationship between the binding of monoclonal antibody (MAb) F7-26 to cellular DNA and the cytotoxicity of nitrogen mustard derivatives and nitrosoureas in the range of 1-6 log cell killing; and the ability of MAb to distinguish between sensitive and drug-resistant cells. The major question to be answered in this study is whether a quantitative correlation between MAb binding and drug sensitivity or resistance can be observed in different murine and human tumor cells. MAb binding to DNA in individual cells will be measured by flow cytometry. The correlation between MAb F7-26 immunoreactivity with DNA in the cells treated with alkylating agents and cell killing will be studied in two paired cell lines sensitive and resistant to alkylating agents: i) lung carcinoma lines A549 (Mer+ phenotype) and A427 (Mer- phenotype) resistant and sensitive to nitrosoureas, respectively; ii) ovarian carcinoma cell lines A2780 sensitive and resistant to L-PAM. In each pair of lines, relationship between cell survival, initial DNA damage and repair of DNA damage after drug removal will be studied. Mouse solid tumor Lewis lung carcinoma and its sublines resistant to nitrosoureas ans cyclophosphamide will be treated with drugs in vitro and in vivo. The binding of MAb to aneuploid tumor cells will be compared with tumor response to treatment. The relationship between the appearance of cell subsets with low DNA immunoreactivity and the development of drug resistance during repeated drug treatments will be studied. To characterize the type of change in DNA detected with the Mab the antigenic determinant of MAb F7-26 will be studied by direct and competitive ELISA immunoassay. The relative affinity of MAb to different deoxyribohomopolymers and the number of bases in oligonucleotides necessary to bind the MAb will be determined. Flow cytometric study of drug effects on DNA in individual tumor cells proposed herein should make it possible to apply the new methodology in clinical settings.