FLOW CYTOMETRIC STUDY OF ANTICANCER DRUG EFFECTS ON DNA

本研究的总体目标是实验和临床 DNA损伤分析在单个细胞中的应用 用抗DNA单克隆抗体对细胞进行染色。 流式细胞术 用烷化剂处理的细胞中DNA损伤的测量 显示出评估异质细胞群体中的耐药性， 分析DNA修复并选择治疗上有用的DNA修复 抑制剂的 DNA修复及其化疗药物的抑制作用 将研究开发筛选系统，以便进行有效的修复 抑制剂，为联合化疗提供合理依据 基于DNA修复抑制。 不同药物的作用 类及其组合的DNA损伤的修复诱导的 代表主要肿瘤定位的细胞系中的烷化剂 将被研究。 临床应用的目标是开发 卵巢癌、多发性骨髓瘤、慢性 骨髓性白血病和慢性淋巴性白血病。 实体瘤将是 在体外用烷化剂处理，分解和DNA损伤 将在非整倍体肿瘤细胞中测量。 从血液中分离的细胞 骨髓会被烷化剂和DNA损伤处理 将在流式细胞仪上测量。 DNA损伤的强度 将体外药物诱导的反应与患者的反应进行比较 到使用烷化剂的化疗。 DNA修复的效果 抑制剂对烷化剂诱导的卵巢肿瘤DNA损伤的作用 将进行体外研究，以确定 个体肿瘤的DNA修复抑制。 DNA修复的速度， 通过DNA免疫反应性的降低来评价白血病细胞 在无药物培养基中后孵育期间。 修复率将是 与耐药性的发展有关。 细胞的作用 将研究耐药性发展中的相互作用， 细胞培养和人类肿瘤的异种移植物。 药物筛选系统 其消除基于细胞亚群与 将产生不同的抗性机制。

The general goal of this study is the experimental and clinical applications of DNA damage assay in individual cells based on the staining of cells with anti-DNA monoclonal antibody. Flow cytometric measurements of DNA damage in the cells treated with alkylating agents was shown to evaluate drug resistance in heterogeneous cell populations, to analyze DNA repair and to select therapeutically useful DNA repair inhibitors. DNA repair and its inhibition by chemotherapeutic agents will be studied to develop screening system for efficient repair inhibitors and to develop rational basis for combination chemotherapy based on DNA repair inhibition. The effects of drugs of different classes and their combinations of the repair of DNA damage induced by alkylating agents in cell lines representing major tumor localizations will be studied. The goal of clinical applications is to develop chemosensitivity tests for ovarian carcinomas, multiple myeloma, chronic myeloid leukemia and chronic lymphoid leukemia. Solid tumors will be treated in vitro with alkylating agents, disaggregated and DNA damage will be measured in aneuploid tumors cells. Cells isolated from blood and bone marrow will be treated with alkylating agents and DNA damage will be measured on a flow-cytometer. The intensity of DNA damage induced by drugs in vitro will be compared with the response of patients to chemotherapy with alkylating agents. The effect of DNA repair inhibitors on DNA damage induced by alkylating agents in ovarian tumors in vitro will be studied in order to determine the sensitivity of individual tumors to DNA repair inhibition. The rate of DNA repair in leukemic cells will be evaluated by decrease of DNA immunoreactivity during post-incubation in drug-free medium. Rate of repair will be correlated with the development of resistance. The role of cell interaction in the development of drug resistance will be studied in cell culture and xenografts of human tumors. Screening system for drugs which eliminate resistance based on interaction of cell subsets with different mechanisms of resistance will be developed.