Davis P, Bazar K, Huper G, Lozano G, Marks J, Iglehart J D
Department of Surgery, Duke University Medical Center, Durham, North Carolina 27710, USA.
Oncogene. 1996 Sep 19;13(6):1315-22.
The p53 gene is a recessive oncogene whose loss of function can result in cell transformation. Approximately 25% of human breast cancers contain missense mutations in one p53 allele, leading to inactivation of the mutated protein. In almost all of these cases, the wild-type allele is also lost. However, it remains uncertain whether mutant p53 acts in a dominant negative fashion over the wild-type protein. Two parameters of p53 function, transcriptional activation and transcriptional repression, were studied under a variety of experimental conditions within malignant and normal breast epithelial cells. Transient transfection of DNA encoding wild-type p53 was able to transactivate p53-responsive promoters. Wild-type p53 functioned equally well in malignant cells which harbored an endogenous mutation in p53, in malignant cells containing normal p53 and in normal mammary epithelial cells. Co-transfection of cDNAs encoding mutant p53 proteins were unable to inhibit the ability of wild-type p53 to transactivate the reporter constructs. Repression of viral promoters by normal p53 protein was not inhibited by endogenous or co-transfected mutant p53 -proteins. Finally, the p53 regulated gene WAF1/CIP1/p21 was induced following gamma irradiation in normal mammary cells, containing endogenous wild-type p53 and in the same cells transfected with mutant p53 genes. From these experiments we conclude that mutant p53 proteins do not inactivate the transactivating (or repressing) function of a co-expressed normal p53 protein in these cells implying that complete loss of wild-type p53 is required to eliminate these functions in breast epithelium.
p53基因是一种隐性癌基因,其功能丧失可导致细胞转化。约25%的人类乳腺癌在一个p53等位基因中存在错义突变,导致突变蛋白失活。在几乎所有这些病例中,野生型等位基因也会丢失。然而,尚不确定突变型p53是否以显性负性方式作用于野生型蛋白。在恶性和正常乳腺上皮细胞的各种实验条件下,研究了p53功能的两个参数,即转录激活和转录抑制。编码野生型p53的DNA瞬时转染能够反式激活p53反应性启动子。野生型p53在携带p53内源性突变的恶性细胞、含有正常p53的恶性细胞和正常乳腺上皮细胞中功能同样良好。共转染编码突变型p53蛋白的cDNA不能抑制野生型p53反式激活报告构建体的能力。正常p53蛋白对病毒启动子的抑制作用不受内源性或共转染的突变型p53蛋白的抑制。最后,在含有内源性野生型p53的正常乳腺细胞以及转染了突变型p53基因的相同细胞中,γ射线照射后诱导了p53调控基因WAF1/CIP1/p21。从这些实验中我们得出结论,在这些细胞中,突变型p53蛋白不会使共表达的正常p53蛋白的反式激活(或抑制)功能失活,这意味着在乳腺上皮中要消除这些功能需要野生型p53完全丧失。