Gerhards R E, Mehnert F E
J Cell Biochem. 1986;31(1):27-39. doi: 10.1002/jcb.240310105.
A rabbit antiserum (A2) directed against the detergent-solubilized fraction of the simian virus 40-transformed mouse embryo fibroblast cell line VLM detects common antigens in primary cell cultures from BALB/c mouse embryos and in transformed cell lines from various species. Positively reacting cell cultures show a set of polypeptides with molecular weight species p86, p74, p68, p46, p42, p40, and p35. As tested by Western blotting procedures, all immunoprecipitated proteins carry immunologically reactive determinants. By analysis with two-dimensional gel electrophoresis, all precipitated polypeptides show charge heterogeneities. Concerning the two major members of the protein set, p40 consists of at least four subspecies with isoelectric points in the range of pH 6.2-6.8, whereas p35 is composed of two subspecies focusing between pH 6.4 and pH 7.2. By comparison of the two-dimensional patterns of p35 of various transformed cell lines, a basic (pH 6.6-7.2) and an acidic (6.4-6.6) charge type of p35 could be observed. Comparative analyses of primary cell cultures from 12-16-day mouse embryos show the immunoprecipitated set of polypeptides only in the 16-day embryo cell cultures. After six further propagations, these cells express the immunoreactive proteins as strongly as the primary cell cultures. In embryonic cell cultures of day 14 of gestation the expression of this set of antigens is induced only when cells are propagated at least six times. Under identical conditions these proteins could not be induced in cell cultures of 18-day-old mouse embryos. None of the polypeptides could be immunoprecipitated from primary mouse kidney cell cultures of 12-day-old mice even when the cultures were propagated at least 15 times. This set of polypeptides is also present in simian virus 40-transformed cells of hamster, rat, monkey, and human origin. These findings suggest that in simian virus 40-transformed mouse cells, in addition to p53, the synthesis of other embryonic antigens is reactivated. The presence of the described set of polypeptides in polyoma virus-transformed cells of rat and mouse origin and in cell lines derived from malignant human tumors might indicate common functions in metabolic patterns of transformed cells.
一种针对猿猴病毒40转化的小鼠胚胎成纤维细胞系VLM去污剂可溶部分的兔抗血清（A2），能在BALB/c小鼠胚胎的原代细胞培养物以及来自各种物种的转化细胞系中检测到共同抗原。呈阳性反应的细胞培养物显示出一组分子量分别为p86、p74、p68、p46、p42、p40和p35的多肽。经蛋白质印迹法检测，所有免疫沉淀的蛋白质都带有免疫反应性决定簇。通过二维凝胶电泳分析，所有沉淀的多肽都显示出电荷异质性。关于该蛋白质组中的两个主要成员，p40至少由四个等电点在pH 6.2 - 6.8范围内的亚类组成，而p35由两个聚焦在pH 6.4和pH 7.2之间的亚类组成。通过比较各种转化细胞系中p35的二维图谱，可观察到p35的一种碱性（pH 6.6 - 7.2）和一种酸性（6.4 - 6.6）电荷类型。对12 - 16天龄小鼠胚胎的原代细胞培养物进行比较分析发现，仅在16天龄胚胎细胞培养物中能免疫沉淀出该组多肽。再传代六次后，这些细胞表达免疫反应性蛋白的强度与原代细胞培养物相同。在妊娠第14天的胚胎细胞培养物中，只有当细胞至少传代六次时，这组抗原的表达才会被诱导。在相同条件下，18天龄小鼠胚胎的细胞培养物中无法诱导出这些蛋白质。即使将12天龄小鼠的原代肾细胞培养物传代至少15次，也无法从其中免疫沉淀出任何一种多肽。这组多肽也存在于仓鼠、大鼠、猴和人源的猿猴病毒40转化细胞中。这些发现表明，在猿猴病毒40转化的小鼠细胞中，除了p53之外，其他胚胎抗原的合成也被重新激活。在大鼠和小鼠源的多瘤病毒转化细胞以及源自人类恶性肿瘤的细胞系中存在所述的这组多肽，这可能表明它们在转化细胞的代谢模式中具有共同功能。
