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p53 transformation-related protein: Detection by monoclonal antibody in mouse and human cells

机译：p53转化相关蛋白：小鼠和人类细胞中单克隆抗体的检测

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A transformation-related protein of Mr 53,000, designated p53, has been detected in a range of neoplastic cell types of the mouse by using immunoprecipitation of [35S]-methionine-labeled cell extracts with mouse antiserum [DeLeo, A. B., Jay, G., Appella, E., DuBois, G. C., Law, L. W. & Old, L. J. (1979) Proc. Natl. Acad. Sci. USA 76, 2420-2424]. We have now prepared a monoclonal antibody to p53 and have used it to study the occurrence and intracellular location of p53 by indirect immunofluorescence assays. In accordance with the results of immunoprecipitation, these tests showed p53 in all 13 transformed mouse cell lines studied. In each case, p53 was found in the nucleus. No p53 was detected in normal mouse fibroblasts, 3T3 cells, bone marrow cells, thymus cells, or embryo cells. A serologically related protein was detected in the nucleus of human cells by monoclonal antibody and was found in both normal and neoplastic cultured cells. Expression of p53 in human cells correlates with the growth characteristics of the culture, high p53 levels being associated with rapid cell proliferation and low p53 levels, with cessation of cell division. Normal and malignant human cells differ, however, with regard to the effect of confluency on p53 expression. Normal kidney epithelium and fetal brain cells, which express high p53 levels during exponential growth, show a prompt decrease in p53 associated with contact inhibition of cell division. Malignant cells, on the other hand, continue to express p53 after confluency and subsequent overgrowth of the monolayers. These results suggest that p53 may be involved in the normal regulation of cell division and that malignant transformation leads to abnormalities in the control of p53 expression.
机译：通过使用[35S]-蛋氨酸标记的细胞提取物与小鼠抗血清[DeLeo，AB，Jay，G.]免疫沉淀，在一系列小鼠的肿瘤细胞类型中检测到了53,000先生的转化相关蛋白p53。 ，Appella，E.，DuBois，GC，Law，LW＆Old，LJ（1979）Proc。 Natl。学院科学USA 76，2420-2424]。现在，我们已经制备了针对p53的单克隆抗体，并已通过间接免疫荧光分析法将其用于研究p53的发生和细胞内位置。根据免疫沉淀的结果，这些测试显示在所有研究的13种转化的小鼠细胞系中都存在p53。在每种情况下，在细胞核中都发现有p53。在正常小鼠成纤维细胞，3T3细胞，骨髓细胞，胸腺细胞或胚胎细胞中未检测到p53。通过单克隆抗体在人细胞核中检测到血清学相关蛋白，并且在正常和肿瘤培养细胞中均发现。人细胞中p53的表达与培养物的生长特性有关，高p53水平与细胞快速增殖有关，而低p53水平则与细胞分裂停止有关。然而，正常细胞和恶性人类细胞在融合对p53表达的影响方面有所不同。正常的肾上皮和胎儿脑细胞在指数增长过程中表达高的p53水平，显示p53的迅速下降与细胞分裂的接触抑制有关。另一方面，在单层融合和随后的过度生长后，恶性细胞继续表达p53。这些结果表明p53可能参与细胞分裂的正常调节，并且恶性转化导致p53表达控制异常。

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