HUMAN NEUROBLASTOMA CELL TRANSDIFFERENTIATION

人神经母细胞瘤细胞转分化

• 批准号: 3182080
• 负责人: JUNE L. BIEDLER
• 金额: $ 13.83万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 1989-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3182080
• 关键词: 3'5' cyclic nucleotide phosphodiesterase; aromatic L aminoacid decarboxylase; athymic mouse; cancer registry /resource; cell adhesion; cell sorting; cell transformation; cell type; clone cells; cytoskeleton; dopamine beta monooxygenase; enzyme mechanism; extracellular matrix; gene expression; growth media; histochemistry /cytochemistry; human tissue; immunochemistry; karyotype; laboratory mouse; monoclonal antibody; natural gene amplification; neoplasm /cancer genetics; neoplasm /cancer pharmacology; neoplastic cell; neuroblastoma; oncogenes; radionuclide double label; surface antigens; tyrosine 3 monooxygenase

Human neuroblastoma cell lines sometimes comprise two distinctly different morphological types of cells. One cell type (termed N) is small, has neuritic processes and adheres poorly to the substrate. The other is large, flat, and highly substrate-adherent (termed S). Karyotype analysis indicates that N and S cells from any one cell line arise from a common precursor. Moreover, as shown for the SK-N-SH cell line, N and S cells can undergo bidirectional morphological and biochemical interconversion. Whereas N cells express enzyme activities of noradrenergic neurons, S cells (of the SK-N-SH line) do not contain these enzymes, but rather, express activity for the melanocyte enzyme tyrosinase. New studies show that N and S cells also can be distinguished by expression of cell surface antigens revealed by serological analysis with a panel of monoclonal antibodies. Further, N and S cells show differential expressions of collagen, neurofilament protein, vimentin, and fibronectin. These results thus indicate that human neuroblastoma cells in culture undergo spontaneous bidirectional interconversion between cellular phenotypes (neuronal, melanocytic, glial) within the neural crest repertoire. This transdifferentiation phenomenon will be explored by continued isolation of N and S clones from a large group of neuroblastoma lines and characterization of clones as to 1) karyotype; 2) marker enzymes of neural crest cell derivatives; 3) cell surface antigen expression (mixed hemadsorption assays with a monoclonal antibody panel and EGF receptor binding assays); and 4) cytoskeletal and extracellular matrix proteins [neurofilaments, vimentin, GFAP, collagen(s), fibronectin]. The mechanisms of interconversion will be studied by assessing changes in the transcription and/or translation of oncogenes N-myc and c-src, EGF receptor, collagen isotypes and neurofilaments. Nude mice will be used to determine tumorigenic and invasive potentials of the two cell types. Differences between N and S cells in response to cancer chemotherapeutic agents will be investigated. The ultimate objective is to assess and exploit the natural phenotypic heterogeneity exhibited by human neuroblastoma cells and to understand the coordinate program of phenotypic transdifferentiation in terms of both malignant transformation and cell differentiation.

人神经母细胞瘤细胞系有时包含两种截然不同的细胞系 细胞的形态类型。 一种细胞类型（称为 N）较小，具有 神经炎过程和对基质的粘附不良。 另一个是 大、平坦且与基材高度粘附（称为 S）。 核型分析 表明来自任何一种细胞系的 N 和 S 细胞来自共同的 前驱。 此外，如 SK-N-SH 细胞系所示，N 和 S 细胞可以 进行双向形态和生化相互转化。 N 细胞表达去甲肾上腺素能神经元的酶活性，S 细胞表达 （SK-N-SH 系的）不包含这些酶，而是表达 黑素细胞酪氨酸酶的活性。 新的研究表明 N 和 S细胞还可以通过细胞表面抗原的表达来区分 通过一组单克隆抗体的血清学分析揭示。 此外，N 和 S 细胞表现出胶原蛋白的差异表达， 神经丝蛋白、波形蛋白和纤连蛋白。 这些结果因此 表明培养中的人神经母细胞瘤细胞自发地发生 细胞表型之间的双向相互转换（神经元、 神经嵴库中的黑素细胞、神经胶质细胞）。 这 将通过持续分离来探索转分化现象 来自一大群神经母细胞瘤系的 N 和 S 克隆以及 克隆的表征： 1) 核型； 2) 神经标记酶 嵴细胞衍生物； 3）细胞表面抗原表达（混合 使用单克隆抗体组和 EGF 受体进行血吸附测定 结合测定）； 4) 细胞骨架和细胞外基质蛋白 [神经丝、波形蛋白、GFAP、胶原蛋白、纤连蛋白]。 机制 将通过评估的变化来研究相互转化 癌基因 N-myc 和 c-src、EGF 的转录和/或翻译 受体、胶原同种型和神经丝。 裸鼠将被用来 确定两种细胞类型的致瘤和侵袭潜力。 N 细胞和 S 细胞对癌症化疗反应的差异 代理商将被调查。 最终目标是评估和 利用人类表现出的自然表型异质性 神经母细胞瘤细胞并了解表型的协调程序 恶性转化和细胞转分化 差异化。