寡核苷酸芯片检测肺癌患者痰标本中p53点突变  

Clinical study on detection of p53 gene hot-spot mutations in sputum sample of lung cancer using oligonucleotide microarray

作　　者：于海建[1] 安耀武[1] 于群[1] 李龙芸[2] 

机构地区：[1]南方医科大学附属花都医院,广东广州510800 [2]北京协和医院,北京1007302

出　　处：《甘肃医药》2009年第2期81-84,共4页Gansu Medical Journal

摘　　要：目的:用寡核苷酸基因芯片方法检测肺癌患者痰标本中p53基因的热点突变,为肺癌的诊断探索快速、准确的基因检测途径。方法:收集肺癌患者痰标本并提取基因组DNA,用荧光标记的引物对DNA进行p53基因Exon5、Exon 7 PCR扩增,然后用PCR扩增产物与基因芯片进行杂交,通过对杂交信号的检测,判断是否发生点突变,并用测序方法对部分阳性结果进行验证。结果:在27例肺癌患者痰标本中,p53基因突变的检出率为25.92%,其中小细胞肺癌的突变率为40%(2/5)、肺鳞癌的突变率为26.6%(4/15)、肺腺癌的突变率为14.28%(1/7),检测到的突变位点有175(2)、248(1)、249(2),对2例阳性标本的测序证实了基因芯片的结果。结论:寡核苷酸基因芯片法可以快速、准确检测出肺癌患者痰标本中p53基因的点突变,今后可用该法对肺癌患者和高危人群的痰标本进行大规模筛选,提高肺癌的早期诊断率。Objective： Part of the hot - spot mutations of p53 gene was screened in sputum sample of lung cancer using oligonucleotide chips in order to explore rapid and accurate method in gene diagnosis for lung cancer. Methods： Sputum sample of lung cancer patients were collected and genomic DNAs were extracted. Exon5 and Exon7 PCR amplification of p53 were performed, then hybridization of the products of PCR amplification and gene chips were performed. Hybridization signal was detected in order to assess hot - spot mutation . Finally sequencing of parts positive sample was performed to confirm the hot - spot mutation. Results： Of all 27 patients, 25.92% （7/27） was detected with p53 mutation. 40% （3/5） mutations happened in small cell lung cancer, 26.6% （4/15） in squamous carcinoma and 14.28% （1/7） in adenocarcinoma. Mutation loci included 175,248,and 249. Sequencing map confirmed the parts of mutation of p53. Conclusion： Oligonucleotide chips can rapidly screen the p53 gene mutation in sputum sample of lung cancer. In the future, we may screen the sputum sample of patients and high - risk persons on a large scale.

关 键 词：肺癌 寡核苷酸基因芯片 P53基因 点突变 

分 类 号：R734.2[医药卫生—肿瘤] Q75[医药卫生—临床医学]