CHROMATIN PROBES FOR DISTINGUISHING MALIGNANT CELLS

本研究的目的是发展和应用细胞化学和 适用于流式细胞术的生物物理探针或技术， 用于分析细胞核和细胞质结构的细胞分选仪 可以区分正常的增殖，正常的分化， 癌前病变和恶性细胞，以及详细描述 细胞周期的各个代谢阶段。 通过提供新的，客观的， 和定量标准，探针也可以应用于 肿瘤的分类和预后价值在监测反应性 转移到治疗上。 开发探针是为了区分 在增殖和静止的肿瘤细胞之间， 在规划化疗方案中的价值。 使用各种荧光染料，以下细胞特征是 测量：（1）总DNA、RNA和蛋白质含量以及RNA/DNA， 全细胞和分离的细胞核的RNA/蛋白质比率;（2）膜 线粒体的潜力;（3）多聚核糖体RNA与核糖体RNA的比值;（4） 双链到单链RNA;和（5）DNA的可及性， 原位插入各种大小和亲和力的探针，以及 外源核酸酶 此外，核染色质的结构是 使用染料组合并利用能量转移进行分析 现象，研究DNA结合探针的去极化，存在和 SH基团的可及性，对类固醇激素的敏感性，以及 原位变性敏感性。 染色质探针的更多基础研究 包括研究天然核酸之间分子相互作用 酸、同核糖聚合物或脱氧核糖聚合物，以及诸如吖啶橙子或 派洛宁Y 不同的组织培养细胞系，循环，静止和 分化细胞系统，人白血病，淋巴瘤，实体瘤，和 使用临床细胞学标本。 （三）

The objective of this study is to develop and apply cytochemical and biophysical probes or techniques applicable to flow cytometry and cell-sorting instruments for analysis of nuclear and cytoplasmic structure that can distinguish between normal proliferating, normal differentiating, premalignant, and malignant cells, as well as characterize in detail various metabolic phases of the cell cycle. By providing new, objective, and quantitative criteria, the probes also could have an application in the classification of tumors and prognostic value in monitoring responsiveness of tumor cells to therapy. The probes are developed to discriminate between proliferating and quiescent tumor cells, which is of potential value in planning chemotherapy protocols. Using a variety of fluorochromes, the following cell features are measured: (1)\total DNA, RNA, and protein content as well as RNA/DNA and RNA/protein ratios of whole cells and isolated nuclei; (2)\membrane potential of mitochondria; (3)\polysomal versus ribosomal RNA; (4)\ratio of double-stranded to single-stranded RNA; and (5)\accessibility of DNA in situ to intercalating probes of various sizes and affinities as well as to exogenous nucleases. In addition, structure of nuclear chromatin is analyzed using combinations of dyes and exploiting the energy transfer phenomenon, studying depolarization of DNA-bound probes, presence and accessibility of SH groups, sensitivity to steroid hormones, and DNA in situ sensitivity to denaturation. More basic studies on chromatin probes include investigations on molecular interactions between natural nucleic acids, homoribo-\or deoxyribo-polymers, and such dyes as acridine orange or pyronin Y. Different tissue culture cell lines, cycling, quiescent and differentiating cell systems, human leukemias, lymphomas, solid tumors, and clinical cytology specimens are used. (3)