Celis J E, Ratz G P, Celis A
Department of Medical Biochemistry, Aarhus University, Denmark.
Leukemia. 1987 Oct;1(10):707-17.
Analysis by means of computerized two-dimensional gel electrophoresis (NEPHGE, IEF) of the [35S]-methionine labeled proteins secreted by normal human MRC-5 fibroblasts revealed 476 polypeptides (258 acidic and 218 basic), many of which appeared as charge trains due to modification. Similar analysis of the proteins secreted by SV40 transformed MRC-5 fibroblasts (MRC-5 V2) showed a striking decrease in the levels of many of these proteins as well as the appearance (or increased synthesis) of 47 polypeptides that were either absent or present in very low amounts in normal cells. Of the major secreted polypeptides whose relative proportion decreased dramatically in the MRC-5 V2 cells, 15 were found to be abundant components of other normal (nontransformed) fibroblasts (W138, Xeroderma pigmentosum cell lines). Low levels of these radioactively labeled polypeptides were observed in transformed human cell lines of fibroblast (W138, SV40, HT1080), epithelial (HeLa, transformed amnion cells (AMA), A431, A459) and myeloid (HL-60) origin. No major secreted polypeptide from MRC-5 V2 cells was synthesized exclusively by the transformed cell lines.
通过计算机二维凝胶电泳（非平衡pH梯度电泳、等电聚焦）分析正常人MRC-5成纤维细胞分泌的[35S] - 甲硫氨酸标记蛋白，发现了476种多肽（258种酸性多肽和218种碱性多肽），其中许多由于修饰而呈现为电荷条带。对SV40转化的MRC-5成纤维细胞（MRC-5 V2）分泌的蛋白质进行类似分析，结果显示许多此类蛋白质的水平显著下降，同时出现了（或合成增加）47种在正常细胞中不存在或含量极低的多肽。在MRC-5 V2细胞中相对比例急剧下降的主要分泌多肽中，有15种被发现是其他正常（未转化）成纤维细胞（W138、色素性干皮病细胞系）的丰富成分。在成纤维细胞（W138、SV40、HT1080）、上皮细胞（HeLa、转化羊膜细胞（AMA）、A431、A459）和髓细胞（HL-60）来源的转化人细胞系中观察到这些放射性标记多肽的水平较低。MRC-5 V2细胞中没有主要分泌多肽是由转化细胞系单独合成的。
