CONSTANT DENATURANT GEL-ELECTROPHORESIS AS A RAPID SCREENING TECHNIQUE FOR P53 MUTATIONS
CONSTANT DENATURANT GEL-ELECTROPHORESIS AS A RAPID SCREENING TECHNIQUE FOR P53 MUTATIONS
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DOI：
10.1073/pnas.88.19.8405
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发表时间：
1991-10-01
影响因子：
11.1
通讯作者：
FRIEND, SH
中科院分区：
文献类型：
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作者：
BORRESEN, AL;HOVIG, E;FRIEND, SH
目前，p53基因突变似乎是人类癌症中最常见的遗传改变。 这些突变可以发生在基因的许多不同区域。 我们已经开发了一种修改的变性梯度凝胶电泳称为“恒定变性凝胶电泳”（CDGE），它提供了一个快速和敏感的方法来筛选的四个保守区域内的p53基因的大多数p53突变已被报道。 CDGE的灵敏度首先用所有四个保守区域中的已知p53突变进行测试。 CDGE技术，然后被用来筛选32个乳腺癌，已通过免疫组化方法分析改变p53蛋白水平，其DNA已被证明有染色体17 p标记的杂合性丢失。 通过免疫组化技术，32例肿瘤中只有6例p53表达升高。 然而，CDGE在32个肿瘤中的11个中检测到p53突变。 进行DNA序列分析以确定所有11个样品中这些突变的核苷酸位置。 pYNZ22或p144D6标记的杂合性丢失与p53位点的杂合性丢失或CDGE检测到的突变无关。 我们的结论是，CDGE是一种快速，有效的技术来筛选p53突变。
At present, mutation of the p53 gene appears to be the most common genetic alteration found in human cancers. These mutations can occur within many different regions of the gene. We have developed a modification of denaturing gradient gel electrophoresis termed "constant denaturant gel electrophoresis" (CDGE), which provides a rapid and sensitive method to screen the four conserved regions within the p53 gene where the majority of p53 mutations have been reported. The sensitivity of CDGE was first tested with known p53 mutations in all four conserved regions. The CDGE technique was then used to screen 32 breast carcinomas that had been analyzed by immunohistochemical methods for altered p53 protein levels and whose DNA had already been shown to have loss of heterozygosity for a chromosome 17p marker. By immunostaining techniques, only 6 of the 32 tumors had elevated p53 expression. However, CDGE detected p53 mutations in 11 of the 32 tumors. DNA sequence analysis was performed to determine the nucleotide positions of these mutations in all 11 samples. Loss of heterozygosity for the pYNZ22 or p144D6 markers did not associate with either the loss of heterozygosity at the p53 locus or the mutations detected by CDGE. We conclude that CDGE is a rapid and effective technique to screen for p53 mutations.