PROBES FOR FLOW CYTOMETRY

该项目的长期目标是开发新的探头， 用于通过流动分析单个细胞或 图像细胞术，适用于肿瘤诊断和/或预后; 开发的方法学将应用于细胞周期的研究， 细胞对抗肿瘤药物的敏感性。具体目标集中在 细胞凋亡和细胞增殖的标记物的发展。 DNA原位链断裂标记方法的建立 采用外源性末端转移酶将继续;该技术 将与细胞增殖分析相结合。细胞增殖性 潜在的将估计两者，分析表达的 细胞周期特异性蛋白，细胞周期蛋白E和B1以及DNA测量 复制的后者将通过一种新的方法来检测， 将新复制DNA的光解与DNA检测相结合， 链断裂。这种方法提供了几个优于 通过5-溴脱氧尿苷抗体的DNA复制的常规检测， 还将测试其测量DNA修复的能力。一 将选择荧光染料的组合， 测量细胞DNA含量。DNA的多变量分析 含量、细胞死亡（凋亡）和增殖（细胞周期蛋白E和B1 表达或DNA复制）将应用于人类的研究中。 白血病，化疗前和化疗期间，以及乳腺癌 癌症;将评估测量参数的预后价值。 预计这种综合分析，特别是涉及 G1和G2细胞周期蛋白的估计，将具有高预后价值， 将在治疗期间使用，以监测肿瘤反应。

The long term objective of this project is the development of new probes, methods and instrumentation for analysis of individual cells by flow or image cytometry, applicable for tumor diagnosis and/or prognosis; the developed methodologies will be applied In studIes of the cell cycle and cell sensitivity to antitumor drugs. The specific aims are focused on development of markers of apoptosis, and of cell proliferation. Development of the method based on DNA in situ strand break labeling that employs exogenous terminal transferase will be continued; the technique will be combined with analysis of cell proliferation. Cell proliferative potential will be estimated by both, the analysis of expression of the cell cycle specific proteins, cyclin E and B1 and measurement of DNA replication. The latter will be detected by a novel approach, which combines photolysis of the newly replicated DNA with the detection of DNA strand breaks. This approach, which offers several advantages over the conventional detection of DNA replication by 5-bromodeoxyuridine antibody, will also be tested with respect to its ability to measure DNA repair. A combination of fluorochromes will be selected that will also allow measurement of cellular DNA content. The multivariate analysis of DNA content, cell death (apoptosis) and proliferation (cyclin E and B1 expression or DNA replication) will be applied in studies of human leukemias, before and during the course of chemotherapy, and in breast cancer; the prognostic value of the parameters measured will be evaluated. it is expected that such a combined analysis, especially involving estimates of the G1 and G2 cyclins, will be of high prognostic value, and will be used during treatment, to monitor tumor response.