MORPHOLOGICAL MECHANISMS OF ORGANELLE FUNCTION AND TRANSFORMATION IN CULTURE CELL

肿瘤转化会在细胞生理上产生许多变化，其中一些 这可以用形态学技术来研究。我们已经使用了 研究与转型相关的三个区域的形态方法：1)我们 已经使用光镜免疫细胞化学来评估单克隆性 适合作为癌症免疫毒素试剂的抗体 化疗。我们采用了光镜免疫细胞化学的方法 在整个组织中，使用冷冻的人体组织、低温恒温器切片和 过氧化物酶标记法。这种标准化的方法已经被用于 评估HB-21(一种抗人转铁蛋白受体的单抗)， 抗TAC(一种抗人IL-2受体的单抗)，以及其他 产生抗人卵巢癌细胞系的单抗。 2)研究了多药耐药(MDR)现象。 细胞和形态检查化疗药物的蓄积 药物柔红霉素。用该药物作为药物的荧光标记物 培养中的细胞积累，细胞核的光度显微镜研究 在单个细胞中显示多药耐药细胞有加速外流 一种依赖能源的机制。这种快速的形态分析已经使 可以评估多种药物对这种外排的影响。 活性，并显示出主要抑制柔红霉素外排的维拉帕米， 奎尼丁和长春花碱。3)我们使用免疫细胞化学方法 在转化的培养细胞中定位一些主要蛋白质。我们有 定位的“P55”，细胞中一种显示甲状腺激素的主要蛋白质 结合活性，以及“p86”，一种存在于细胞内的热休克蛋白 在许多细胞中大量存在，并被鉴定为肿瘤特异性的 动物系统中的移植抗原。在其他研究中，初步的 使用单细胞显微注射方法的实验表明， 特异性抑制表皮生长因子受体的合成 注射EGF受体基因的反义RNA。

Neoplastic transformation produces many changes in cell physiology, some of which can be studied using morphologic techniques. We have used morphologic methods to study three transformation-related areas: 1) We have employed light microscopic immunocytochemistry to evaluate monoclonal antibodies for suitability as immunotoxin reagents for cancer chemotherapy. We adapted methods for light microscopic immunocytochemistry in whole tissues, using frozen human tissues, cryostat sections and peroxidase labeling methods. This standardized method has been used to evaluate HB-21 (a monoclonal antibody to the human transferrin receptor), anti-TAC (a monoclonal antibody to the human IL-2 receptor), and other monoclonal antibodies generated against human ovarian cancer cell lines. 2) We have studied the phenomenon of multidrug-resistance (MDR) in cultured cells and morphologically examined the accumulation of the chemotherapeutic drug daunomycin. Using this drug as a fluorescence marker of drug accumulation in cells in culture, photometric microscopy studies of nuclei in individual cells showed that MDR cells have an accelerated efflux mechanism that is energy-dependent. This rapid morphologic assay has made it possible to evaluate multiple drugs for their effects on this efflux activity, and has shown major inhibition of daunomycin efflux by verapamil, quinidine, and vinblastine. 3) We have used immunocytochemical methods to localize some major proteins in transformed cultured cells. We have localized "p55", a major protein in cells that shows thyroid hormone binding activity, and "p86", a cytosolic heat-shock protein present in large amounts in many cells and identified as a tumor-specific transplantation antigen in animal systems. In other studies, preliminary experiments using single cell microinjection methods have demonstrated a specific inhibition of epidermal growth factor receptor synthesis following injection of anti-sense RNA for the EGF receptor gene.