用非同位素PCR－SSCP方法检测石蜡包埋乳腺癌p53基因点突变  

Detection of point mutation of p53 gene by rion-isotopic PCR-SSCP in paraffin-embedded breast cancer tissue

作　　者：陈艺华[1] 董敬朋 张素娟 蔡俊杰 陆药丹 

机构地区：[1]广州第一军医大学病理学教研室,南方医院病理科

出　　处：《中华病理学杂志》1994年第6期327-329,T075,共4页Chinese Journal of Pathology

摘　　要：应用一非同位素ＰＣＲ－ＳＳＣＰ方法检测常规石蜡包埋乳腺癌组织中ｐ５３基因点突变。结果显示６０例乳腺癌中，１４例有异常电泳带，表明这些病例相应ＤＮＡ片段中发生了点突变，其中４例位于第５～６外显子，３例位于第７外显子，７例位于第８外显子。免疫组化法检测突变型ｐ５３蛋白的表达，１３例为阳性，其中１１例ＳＳＣＰ检测到异常电泳带，另２例突变可能发生在所检测的基因片段以外。而５例ＳＳＣＰ分析发现基因突变者，免疫染色却为阴性，主要可能由于石蜡包埋组织中ｐ５３抗原性减弱或消失所致，因此，ＳＳＣＰ法检测ｐ５３基因点突变更为准确、可靠，而且可初步将突变定位在基因某一片段。本研究结果表明非同位素ＰＣＲ－ＳＳＣＰ是一简便、快速、有效的检测基因点突变的方法，特别适用于对大量标本基因点突变的筛选；ｐ５３基因点突变与突变型ｐ５３蛋白免疫染色阳性呈相关关系，但并不完全吻合。on-isotopic PCR-SSCP method was used to detect point mutation of p53 gene in paraffin-em-bedded breast cancers. The abnormal shifting of the single-stranded DNA(ssDNA) was identified in 14 out of 60 cases. The point mutations of four cases were found in exons 5～6,three in exon 7 and sev-en in exon 8. The mutant p5 3 protein was detected by immunohistochemistry, The positive nuclear staining was observed in 13 cases, among which 11 cases showed abnormal shifting of ssDNA in SSCP analysis.The point mutations of the other two cases might occur in the regions which were not exam-ined in this study. The point mutations of p53 were found in 5 cases in the absence of detectable protein.This probably resuIted from the reduction or loss of antigenecity of p53 in some of the routinely pro cessed tissues.Our resuIts indicate that detection of point mutations of p53 gene by PCR-SSCP method is more accurate and reliable than that by immunohistochemistry and non-isotopic PCR-SSCP is espe-cially useful in the screening of point mutations during handling a large number of samples.

关 键 词：乳腺肿瘤 基因P53 突变 聚合酶链反应 

分 类 号：R737.904[医药卫生—肿瘤]