ANALYSIS OF POLYPEPTIDE CHANGES DURING CELLULAR DIFFERENTIATION

该项目的发起是为了定性和 从数量上讲，细胞总蛋白模式在细胞内 运用数量化技术进行辨证转化 双向电泳法。到目前为止取得的成果包括：1) 小鼠表皮细胞在低钙或低钙的情况下培养。 (0.02-0.1毫米)(主要是基底细胞)或高钙(1.2毫米)(主要是成熟细胞) 角质形成细胞)，然后用12-0-十四酰-13-乙酸酯(TPA)处理 1小时、4小时或24小时。表皮基底层细胞的分化 将培养液中的Ca+2从0.05 mM增加到1.2 mM或通过 用TPA治疗。表皮多肽的二维分析 电泳法分析表明，合成的8种多肽分别为 增加，六个减少，至少一个的磷酸化 Ca+2和TPA对多肽的影响相似。这些常见的多肽 与Ca+2和TPA诱导的分化有关的变化可能 涉及佛波酯受体、蛋白激酶C。2)分析 正常支气管上皮细胞(NHBE)多肽的变化及意义 Harvey v-ras转染系、TbE-1、TbE-1SA和TbE-1SAT显示 4个新的多肽(等电点5.5-5.75/56-55 kDa； 4.9/55 kDa；6.4/66 kDa；6.7/39 kDa)从NHBE到最恶性 TBE-1SAT线。还观察到了许多数量上的变化，大多数 值得注意的是，两个多肽(5.6/40 kDa)的表达逐渐丧失 和5.7/40 kDa)，在TBE-1SAT细胞中几乎完全不存在。3) 亚硫酸氢钠转化的仓鼠胚胎细胞的特征 两个多肽1和2(等电点5.1/52 kDa和5.1/28 kDa)的位移， 两个新的多肽3和4(6.8/44 kDa和5.5/46 kDa)的表达 缺失1个多肽59个(6.0/55 kDa)。早期多肽变化 对一系列20个亚硫酸盐转化株系进行了分析。多肽 1、2和5中的更改与 转化过程，并与形态变化有关，而 多肽3和4似乎发生得更晚，而且联系更紧密 随着肿瘤致瘤性的获得。

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. Results obtained to date include: 1) Mouse epidermal cells were cultured in the presence of either low calcium (0.02-0.1 mM) (mainly basal cells) or high calcium (1.2 mM) (mainly mature keratinocytes) and then treated with 12-0-tetradecanoyl-13-acetate (TPA) for 1, 4 or 24 hours. Differentiation of epidermal basal cells were induced by increasing Ca+2 from 0.05 to 1.2 mM in the medium or by treatment with TPA. Analysis of epidermal polypeptides by 2-D electrophoresis indicated that the synthesis of eight polypeptides were increased and six decreased, and the phosphorylation of at least one polypeptide is altered similarly by Ca+2 and TPA. These common polypeptide changes, both associated with differentiation induced by Ca+2 and TPA, may involve the phorbol ester receptor, protein kinase C. 2) Analysis of polypeptide changes from normal bronchial epithelial cells (NHBE) and the Harvey v-ras transfected lines, TBE-1, TBE-1SA, and TBE-1SAT revealed the progressive appearance of four new polypeptides (pI 5.5-5.75/56-55 kDa; 4.9/55 kDa; 6.4/66 kDa; and 6.7/39 kDa) from NHBE to the most malignant TBE-1SAT line. Numerous quantitative changes were also observed, most notably, the progressive loss of expression of two polypeptides (5.6/40 kDa and 5.7/40 kDa), which were almost completely absent in TBE-1SAT cells. 3) Sodium bisulfite transformed hamster fetal cells were characterized by the shift of two polypeptides 1 and 2 (pI 5.1/52 kDa and 5.1/28 kDa), the expression of two new polypeptides 3 and 4 (6.8/44 kDa and 5.5/46 kDa) and the loss of one polypeptide 5 9(6.0/55 kDa). Early polypeptide changes were analyzed in a series of 20 bisulfite transformed lines. Polypeptide changes in 1, 2, and 5 were associated with early steps in the transformation process and related to morphologic changes, whereas polypeptides 3 and 4 appear to occur later and are more closely associated with the acquisition of tumorigenicity.