Molès J P, Moyret C, Guillot B, Jeanteur P, Guilhou J J, Theillet C, Basset-Sèguin N
Laboratoire de Dermatologie Moléculaire, CNRS/CRBM BP 5051, Montpellier, France.
Oncogene. 1993 Mar;8(3):583-8.
In the present study we analysed 38 epithelial skin cancers, 19 basal cell carcinomas (BCCs), 13 squamous cell carcinomas (SCCs) and six Bowen diseases (BwDs), using a combination of polymerase chain reaction (PCR) and single-stranded conformation polymorphism (SSCP) techniques for the presence of p53 and RAS gene mutations. Whereas 48% (9/19) of the BCCs tested presented a mutated p53 gene, the frequency was lower (15%, 2/13) in our series of SCCs and negative in the BwDs. Nine of the 11 characterized mutations were single-nucleotide substitutions and, interestingly, seven of these involved CC dimers, where a C was changed into a T or a G (three C-->T transitions and four C-->G transversions). This mutational pattern, added to the fact that all the mutated tumors occurred at sun-exposed body sites, implicates UV light in their genesis. Furthermore, we observed two internal deletions of 6 and 24 bp whose flanking sequences contained two or three Cs on either strand. In addition to molecular detection, we searched for p53 protein accumulation, by immunocytochemical staining, in a subset of 23 epithelial skin tumors (nine bearing a mutation, 14 which scored negative in our assay). Three commercially available anti-p53 antibodies (PAb CM1, mAbs DO7 and 1801) were used, and 3/23 (all showing a mutated p53 gene) presented specific nuclear staining. In contrast to other reported data we could not detect any activating RAS gene mutation in our series of human skin cancers.
在本研究中,我们分析了38例上皮性皮肤癌,其中包括19例基底细胞癌(BCC)、13例鳞状细胞癌(SCC)和6例鲍温病(BwD),采用聚合酶链反应(PCR)和单链构象多态性(SSCP)技术相结合的方法检测p53和RAS基因突变的存在。在所检测的BCC中,48%(9/19)呈现p53基因突变,而在我们的SCC系列中,该频率较低(15%,2/13),在BwD中则为阴性。11个已鉴定的突变中有9个是单核苷酸替换,有趣的是,其中7个涉及CC二聚体,即一个C被替换为T或G(3个C→T转换和4个C→G颠换)。这种突变模式,再加上所有突变肿瘤均发生在阳光暴露的身体部位这一事实,提示紫外线与它们的发生有关。此外,我们观察到两个分别为6 bp和24 bp的内部缺失,其侧翼序列在两条链上均含有两个或三个C。除了分子检测外,我们还通过免疫细胞化学染色在23例上皮性皮肤肿瘤的一个亚组中寻找p53蛋白的积累情况(9例有突变,14例在我们的检测中为阴性)。使用了三种市售的抗p53抗体(PAb CM1、mAb DO7和1801),23例中有3例(均显示p53基因突变)呈现特异性核染色。与其他报道的数据相反,在我们的人类皮肤癌系列中未检测到任何激活的RAS基因突变。