石蜡包埋的石棉肺癌组织中p53基因突变的分析方法

Detection of p53 Gene Mutation in Paraffin embedded Asbestos related Lung Cancer Tissue

为了利用石蜡包埋的病理组织来研究石棉肺癌的基因突变情况，选用１０例石蜡包埋的石棉肺癌组织进行ＰＣＲ－ＳＳＣＰ分析，检测其抗癌基因ｐ５３的第５、第７和第８外显子的突变情况。经银染检查，发现４个病例的ｐ５３基因的第７或第８外显子片段呈突变阳性。用放射性自显影的ＰＣＲ－ＳＳ－ＣＰ方法分析这１０例病例，检测到７个病例的ｐ５３基因第７或第８外显子片段呈突变阳性。用两种方法检测均未发现这１０个病例的第５外显子呈突变阳性。银染ＰＣＲ－ＳＳＣＰ检测可检出放射性自显影ＰＣＲ－ＳＳＣＰ分析结果的６０％，二者的结果相符合。因而，简便、灵敏而且危害性小的银染检测方法，可以代替放射性自显影用于ＰＣＲ－ＳＳＣＰ分析，研究石棉肺癌的基因突变情况。

Paraffin embedded asbestos related lung cancer tissue of ten cases was analyzed with polymerase chain reaction single strand conformation polymorphism (PCR SSCP) to study its genetic mutation of exon 5, 7 and 8 of anti oncogene p53 by silver stain. Results showed that fragment of exon 7 or 8 of p53 gene in four cases was positive in silver staining analysis of PCR SSCP. Exon 7 or 8 fragments of p53 gene was detected positive for mutation in seven of these ten cases with autoradiographic analysis of PCR SSCP. No exon 5 fragment was found positive for mutation in these ten cases with both methods. Agreement between results of silver staining and those of autoradiographic PCR SSCP was 60%. Hence, autoradiographic method could be replaced by silver staining, a simpler and more sensitive one, in PCR SSCP to study gene mutation of asbestosrelated lung cancer.