ANALYSIS OF POLYPEPTIDE CHANGES DURING CELLULAR DIFFERENTIATION

该项目的启动是为了分析，无论是定性和 定量地，在细胞生长过程中总细胞蛋白质模式的变化 利用定量技术进行分化和转化， 双向电泳 一种细胞培养系统已经被开发出来 用于研究原发性正常人支气管的分化 上皮（NHBE）细胞通过12-0-十四酰基-13-乙酸酯（TPA）或 转化生长因子β（TGF-β）。 双向凝胶 [14-C]-标记多肽的电泳（2D-PAGE）， 用以下物质处理NHBE细胞和人肺癌细胞系A1146 TPA和TGF-β仅导致定量多肽差异。 NHBE细胞表现出更明显的多肽表达调节， 与A1146相比，用TPA（10 nM）和TGF-β（30 pg/ml）处理后 细胞，TPA诱导分化难治的细胞系， TGF-β 用TPA和TGF-β处理NHBE细胞24小时后，15 多肽特异性地由TPA调节，4个被TPA调节。 单独TGF-β，6人受到TPA和TGF-β的影响。 延长 暴露（72小时）主要导致（38/42）TPA诱导的变化， 只有一种多肽被TGF-β特异性调节。 的42 在72小时时有4条多肽（5.85/88 kDa; 6.10/40 kDa; 5.00/40 kDa;和5.80/28 kDa）也表现出显著性差异。 24小时处理后的定量调节。

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.