Moro F, Ottaggio L, Bonatti S, Simili M, Miele M, Bozzo S, Abbondandolo A
CSTA-Mutagenesis Laboratory, National Institute for Research on Cancer, Genoa, Italy.
Carcinogenesis. 1995 Oct;16(10):2435-40. doi: 10.1093/carcin/16.10.2435.
To answer the question whether the level of p53 expression also reflects the status of a cell, with reference to transformation and genome stability, we have examined, by immunocytochemistry, the presence of p53 protein in a number of cell types including human diploid cells, Chinese hamster embryonal cells at different passages and gene amplified and/or transformed Chinese hamster cell lines. Primary human fibroblasts at early passage (LEO) and an established, non transformed, Chinese hamster cell line at early passage (CHEF/18) did not show any detectable p53 expression, either nuclear or cytoplasmic. All transformed human (Raji) and Chinese hamster cell lines (CHO, V79, V79/B7) showed a nuclear expression of p53, although at different intensities. Two cell lines selected from V79/B7 for their resistance to phosphonacetyl-L-aspartate or methotrexate and previously shown to bear gene amplification, showed p53 expression. In PALA L cells p53 expression was nuclear as in other positive cell lines tested, while in MTX M cells it was cytoplasmic. CHEF/18 cells at late passage in culture showed the typical behaviour of transformed cells and p53 was detected in several cells. Moreover, when transformed CHO cells were treated with compounds known to induce reverse transformation, both the disappearance of hallmarks of transformed phenotype and p53 reduction were observed. These results indicate a strong association within the same cell type between p53 expression and transformed status.
为了回答p53表达水平是否也反映细胞状态这一问题,我们参考细胞转化和基因组稳定性,通过免疫细胞化学检测了多种细胞类型中p53蛋白的存在情况,这些细胞类型包括人二倍体细胞、不同传代阶段的中国仓鼠胚胎细胞以及基因扩增和/或转化的中国仓鼠细胞系。早期传代的原代人成纤维细胞(LEO)和早期传代的已建立的未转化中国仓鼠细胞系(CHEF/18),无论在细胞核还是细胞质中均未显示出任何可检测到 的p53表达。所有转化的人细胞系(Raji)和中国仓鼠细胞系(CHO、V79、V79/B7)均显示出p53的细胞核表达,尽管强度不同。从V79/B7中选出的两个对膦酰乙酰-L-天冬氨酸或甲氨蝶呤具有抗性且先前已证明存在基因扩增的细胞系,显示出p53表达。在PALA L细胞中,p53表达与其他测试的阳性细胞系一样位于细胞核中,而在MTX M细胞中则位于细胞质中。培养后期传代的CHEF/18细胞表现出转化细胞的典型行为,并且在一些细胞中检测到了p53。此外,当用已知可诱导反向转化的化合物处理转化的CHO细胞时,观察到转化表型特征消失且p53减少。这些结果表明在同一细胞类型中p53表达与转化状态之间存在很强的关联。