ANALYSIS OF POLYPEPTIDE CHANGES DURING CELLULAR DIFFERENTIATION

细胞分化过程中多肽变化的分析

• 批准号: 3963492
• 负责人: P J WIRTH
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 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/3963492
• 关键词: cell differentiation; chemical carcinogenesis; gel electrophoresis; gene expression; growth media; lung neoplasms; respiratory epithelium; tissue /cell culture; transforming growth factors; unsaturated fatty acids

This project was initiated to analyze, both qualitatively and quantitatively, changes in total cellular protein patterns during cellular differentiation and transformation using the technique of quantitative two-dimensional electrophoresis. A cell culture system has been developed for the study of differentiation of primary normal human bronchial epithelial (NHBE) cells by 12-0-tetradecanoyl-13-acetate (TPA) or transforming growth factor type beta (TGF-beta). Two-dimensional gel electrophoresis (2D-PAGE) of [14-C]-labeled polypeptides following treatment of NHBE cells and the human lung carcinoma cell line, A1146, with TPA and TGF-beta resulted in only quantitative polypeptide differences. NHBE cells showed a more pronounced modulation of polypeptide expression following treatment with TPA (10 nM) and TGF-beta (30 pg/ml) than A1146 cells, a cell line refractory to differentiation induction by TPA and TGF-beta. After 24 hr treatment of NHBE cells with TPA and TGF-beta, 15 polypeptides were specifically modulated by TPA, 4 were modulated by TGF-beta alone, and 6 were affected by both TPA and TGF-beta. Prolonged exposure (72 hr) resulted in predominantly (38/42) TPA-induced changes and only one polypeptide was specifically modulated by TGF-beta. Of the 42 polypeptides significantly modulated at 72 hr, 4 polypeptides (5.85/88 kDa; 6.10/40 kDa; 5.00/40 kDa; and 5.80/28 kDa) also exhibited significant quantitative modulation after 24 hr treatment.

该项目的启动是为了进行定性和分析 定量地，细胞过程中总细胞蛋白质模式的变化 使用定量技术进行分化和转化 二维电泳。 已开发出细胞培养系统 用于研究原发性正常人支气管的分化 12-0-十四烷酰基-13-乙酸酯 (TPA) 或 β 型转化生长因子（TGF-β）。 二维凝胶 [14-C]标记多肽的电泳 (2D-PAGE) NHBE细胞和人肺癌细胞系A1146的处理 TPA和TGF-β仅导致多肽数量上的差异。 NHBE 细胞表现出更明显的多肽表达调节 与 A1146 相比，使用 TPA (10 nM) 和 TGF-β (30 pg/ml) 治疗后 细胞，一种难以通过 TPA 诱导分化的细胞系， 转化生长因子-β。 用 TPA 和 TGF-β 处理 NHBE 细胞 24 小时后，15 多肽受 TPA 特异性调节，其中 4 个受 TPA 调节 仅 TGF-β，6 人同时受到 TPA 和 TGF-β 的影响。 延长 暴露（72小时）主要导致（38/42）TPA引起的变化， 只有一种多肽被TGF-β特异性调节。 42 人中 72 小时显着调节的多肽，4 种多肽（5.85/88 kDa； 6.10/40 kDa； 5.00/40 kDa；和 5.80/28 kDa）也表现出显着的 24小时处理后进行定量调节。