MORPHOLOGICAL MECHANISMS OF ORGANELLE FUNCTION AND TRANSFORMATION IN CULTURE CELL

肿瘤转化会引起细胞的许多变化 生理学，其中一些可以用形态学来研究 技巧。我们用形态学方法研究了三个 一般领域：1)我们使用了光学显微镜 免疫细胞化学法评价和筛选单抗 用于癌症治疗的免疫毒素试剂的抗体。一个 设计了大规模的筛选室，用于 免疫荧光作为表面反应的首选方法 抗人卵巢癌细胞的单抗。杂交瘤 用这种方法选择的克隆通过筛选进一步评估 过氧化物酶抗人正常组织和卵巢肿瘤的研究 冰冻切片中的免疫细胞化学。使用这些方法 我们发现了两个与人卵巢高度反应的杂交瘤。 肿瘤：OVB1和OVB3。我们已经确定了组织的特征 这些抗原在人体内的分布和生化特性 在肿瘤和正常组织中培养的细胞。2)我们有 对青壮年多药耐药(MDR)现象进行研究 培养的细胞，并使用单抗(MRK-16)来 以人mdrl基因产物(P170)为探针演示 耐药KB表面和高尔基体中的p170 培养中的细胞。我们还显示涂层中没有P170 PITS，表明这种蛋白被固定在血浆中 薄膜。我们还用这种抗体检测了 P170在正常人体组织中的分布，我们有 评估P170在渐进性耐药中的数量 使用免疫荧光的KB细胞的衍生物。3)在其他 研究表明，我们已经定位了P55，一种主要的甲状腺激素结合 内质网和核膜中的蛋白质 用免疫过氧化物酶检测正常组织中的培养细胞 组织化学。这种蛋白质在细胞中的含量最高。 有丰富的内质网。其他细胞化学物质 研究表明，笼状蛋白包被的凹坑在减少 血管紧张素转换酶在离体兔体内的水转运 皮质集合管。我们还演示了本地化 与小麦胚凝集素反应的碳水化合物在细胞核上的位置 孤立核中的孔洞。

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 general areas: 1) We have employed light microscopic immunocytochemistry to evaluate and select monoclonal antibodies for use as immunotoxin reagents for cancer therapy. A large scale screening chamber has been devised for use with immunofluorescence as a primary selection for surface-reactive monoclonal antibodies to human ovarian cancer cells. Hybridoma clones selected in this way were further evaluated by screening against normal human tissues and ovarian tumors using peroxidase immunocytochemistry in cryostat sections. Using these methods we have found two hybridomas highly reactive with human ovarian tumors: OVB1 and OVB3. We have characterized the tissue distribution and biochemical properties of these antigens in cultured cells, in tumors and in normal tissues. 2) We have studied the phenomenon of multidrug-resistance (MDR) in cultured cells, and have used a monoclonal antibody (MRK-16) to the human mdrl gene product (P170) as a probe to demonstrate P170 on the surface and in the Golgi stacks of drug-resistant KB cells in culture. We also showed the absence of P170 from coated pits, suggesting that this protein is immobilized in the plasma membrane. We have also used this antibody to examine the distribution of P170 in normal human tissues, and we have evaluated the amounts of P170 in progressively resistant derivatives of KB cells using immunofluorescence. 3) In other studies, we have localized p55, a major thyroid hormone binding protein found in the endoplasmic reticulum and nuclear envelope of cultured cells in normal tissues using immunoperoxidase histochemistry. This protein was found in highest amounts in cells that have abundant endoplasmic reticulum. Other cytochemical studies demonstrated a role for clathrin-coated pits in the loss of ADH-responsive water transport in isolated perfused rabbit cortical collecting ducts. We also demonstrated the localization of carbohydrate sites reactive with wheat germ lectin on nuclear pores in isolated nuclei.