Benchimol S, Pim D, Crawford L
EMBO J. 1982;1(9):1055-62. doi: 10.1002/j.1460-2075.1982.tb01296.x.
We have developed quantitative radioimmunological solid phase assays for the host protein p53 from mouse cells and from human cells. The first assay, for mouse p53, depends on having two monoclonal antibodies reacting with different determinants on the p53 molecule. With this assay we have shown that SV40-transformed cells have approximately 100-fold more p53 than untransformed mouse cells and that other transformed cells have intermediate levels. Embryonal carcinoma cell lines have approximately 50-fold less p53 than SV40-transformed cells. This is in contrast to the high levels of incorporation of [35S]methionine into p53 in these cells and indicates that metabolic labelling is not a valid approach for measuring p53 levels. The second assay, for human p53, required a different approach and made use of the anti-p53 antibodies detected in the sera of some breast cancer patients. Human tumour cell lines contained amounts of p53 varying from the high level seen in SV40-transformed human fibroblasts down to less than one hundredth of this amount. Normal human cells showed low levels of p53. The data confirm that many, but not all, human tumour cell lines contain more p53 than normal cells.
我们已经开发出用于检测小鼠细胞和人类细胞中宿主蛋白p53的定量放射免疫固相分析方法。第一种分析方法用于检测小鼠p53,它依赖于两种与p53分子上不同决定簇发生反应的单克隆抗体。通过这种分析方法,我们发现SV40转化细胞中的p53含量比未转化的小鼠细胞多大约100倍,而其他转化细胞中的p53含量处于中间水平。胚胎癌细胞系中的p53含量比SV40转化细胞少大约50倍。这与这些细胞中[35S]甲硫氨酸大量掺入p53的情况形成对比,表明代谢标记法不是测量p53水平的有效方法。第二种分析方法用于检测人类p53,需要采用不同的方法,并利用在一些乳腺癌患者血清中检测到的抗p53抗体。人类肿瘤细胞系中p53的含量各不相同,从SV40转化的人类成纤维细胞中所见的高水平到低于该水平的百分之一。正常人类细胞显示出低水平的p53。这些数据证实,许多(但不是全部)人类肿瘤细胞系中的p53含量比正常细胞多。