Yaginuma Y, Westphal H
Laboratory of Mammalian Genes and Development, National Institute of Child Health and Human Development, NIH, Bethesda, Maryland 20892.
Cancer Res. 1992 Aug 1;52(15):4196-9.
In an effort to analyze molecular mechanisms of human ovarian carcinogenesis, we studied the structure and expression of the p53 gene in different cell lines established from human ovarian carcinomas. In all six lines (PA-1, Caov-3 and -4, OVCAR-3, SK-OV-3, and Kuramochi), p53 abnormalities were detected. In the SK-OV-3 cell line, Southern analysis suggested the presence of sequence deletions/rearrangements in at least one allele of the p53 gene, and transcripts were not detectable by either Northern or polymerase chain reaction analysis. Sequence analysis of the entire coding region of the p53 gene revealed point mutations resulting in codon changes of a highly conserved region of the protein in four cell lines, Caov-3 and -4, OVCAR-3, and Kuramochi. In the Caov-3 cell line, the point mutation resulted in chain termination at codon 136. Quantitation of p53 protein by immunoprecipitation analysis revealed a 6-fold higher than control cell level in PA-1. By contrast, p53 protein was not detectable in lines Caov-3 and SK-OV-3. We conclude that altered levels of p53 gene expression and/or mutant forms of the p53 gene product are associated with all human ovarian cancer cells tested.
为了分析人类卵巢癌发生的分子机制,我们研究了从人类卵巢癌建立的不同细胞系中p53基因的结构和表达。在所有六个细胞系(PA-1、Caov-3和-4、OVCAR-3、SK-OV-3和仓持)中,均检测到p53异常。在SK-OV-3细胞系中,Southern分析表明p53基因至少一个等位基因存在序列缺失/重排,Northern或聚合酶链反应分析均未检测到转录本。对p53基因整个编码区的序列分析显示,在四个细胞系Caov-3和-4、OVCAR-3和仓持中存在点突变,导致该蛋白高度保守区域的密码子发生变化。在Caov-3细胞系中,点突变导致密码子136处的链终止。通过免疫沉淀分析对p53蛋白进行定量,结果显示PA-1中的p53蛋白水平比对照细胞高6倍。相比之下,在Caov-3和SK-OV-3细胞系中未检测到p53蛋白。我们得出结论,p53基因表达水平的改变和/或p53基因产物的突变形式与所有测试的人类卵巢癌细胞相关。