MORPHOLOGICAL MECHANISMS OF ORGANELLE FUNCTION AND TRANSFORMATION IN CULTURE CELL

培养细胞中细胞器功能和转化的形态机制

• 批准号: 3808514
• 负责人: M C WILLINGHAM
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3808514
• 关键词: Pseudomonas; antitumor antibody; cell mediated cytotoxicity; cytoplasm; diagnosis design /evaluation; drug resistance; epidermal growth factor; exotoxins; fluorescent dye /probe; human therapy evaluation; human tissue; immunofluorescence technique; monoclonal antibody; morphology; neoplasm /cancer diagnosis; neoplasm /cancer immunodiagnosis; neoplastic transformation; organelles; ovary neoplasms; protein purification; pyruvate kinase; receptor expression; surface antigens; thyroid hormones; tissue /cell culture; toxin; tumor antigens

There are many changes in the properties of cells following malignant transformation. We have concentrated on changes in the expression of surface molecules that might represent tumor- specific markers that could be exploited both for diagnosis and therapy of human tumors. Using hybridoma technology and morphologic methods, we have isolated monoclonal antibodies to surface epitopes on human tumor cells. One of these antibodies, B3, has recently been studied in detail. This antibody has been shown to react with a specific area of the Lewis(Y) antigen structure. This antibody reacts with some normal tissues, such as gastric and bladder epithelium, as well as human tumors of colon, ovarian and breast origin. Recently, a conjugate of B3 with Pseudomonas exotoxin (PE) has been shown to be effective in killing target tumor cells in tissue culture, as well as in an animal model in nude mice using both A431 and MCF-7 human tumor cells. The absence of serious tissue-specific toxicity in preclinical testing in monkeys suggest that B3 may be useful for therapy of some human cancers. Another antibody, Kl, reactive with ovarian tumors and normal mesothelium has been characterized. It reacts with an antigen that is coexpressed frequently with the CA125 antigen, but it does not react with purified CA125. The antigen reactive with KI (CAK1) is not shed from the surface of cells, yet it is released from cells treated with phospholipase C. CAK1 has been partially purified and appears to be a protein of approximately 40 kD. An immunotoxin prepared using Kl is effective in killing target cells in tissue culture. In other studies, a morphologic assay for the monomer-tetramer conversion of pyruvate kinase Ml in intact cells was developed; this enzyme is a major binding protein for thyroid hormone in the cytoplasm of cells. Also, other studies of a transgenic mouse line developed using the human EGF receptor gene showed that these mice have a defect in microtubule stability in the axoneme of sperm in homozygous animals.

恶性肿瘤发生后，细胞的性质发生了许多变化， 转型 我们已经集中在表达的变化， 可能代表肿瘤特异性标志物的表面分子， 用于诊断和治疗人类肿瘤。 使用 利用杂交瘤技术和形态学方法， 针对人肿瘤细胞表面表位的抗体。 其中一 抗体B3，最近已经详细研究。 该抗体具有 显示与刘易斯（Y）抗原的特定区域反应 结构 这种抗体与一些正常组织反应，如 胃和膀胱上皮，以及结肠、卵巢 和乳房起源。 最近，B3与假单胞菌外毒素的结合物， (PE)已经显示出有效地杀死靶肿瘤细胞， 组织培养，以及在裸鼠的动物模型中使用这两种方法， A431和MCF-7人肿瘤细胞。 缺乏严重的组织特异性 在猴子的临床前试验中的毒性表明，B3可能是有用的 用于治疗某些人类癌症。 另一种抗体K1，与 卵巢肿瘤和正常间皮瘤的特征。 它的反应 使用与CA 125抗原频繁共表达的抗原， 但它不与纯化的CA 125反应。 KI反应性抗原 （CAK 1）不会从细胞表面脱落，但它会从细胞表面释放出来。 用磷脂酶C处理的细胞。CAK 1已部分纯化， 似乎是一种约40 kD的蛋白质。 一种免疫毒素， 在组织培养中使用K1可有效杀死靶细胞。 在 在其他研究中，使用形态学方法测定了 完整细胞中的丙酮酸激酶Ml被开发出来;这种酶是一种主要的 细胞质中甲状腺激素的结合蛋白。 还有， 使用人EGF开发的转基因小鼠系的其他研究 受体基因显示，这些小鼠有缺陷的微管 纯合子动物精子轴丝的稳定性。