High Resolution Melting Analysis for Detecting p53 Gene Mutations in Patients with Non-small Cell Lung Cancer

• Published in: Zhongguo fei ai za zhi 2011-10, Vol.14 (10)
• Main Authors: CHEN, Zhihong, AN, Shejuan, XIE, Zhi, YAN, Honghong, CHEN, Jianguang, SU, Jian, ZHANG, Xuchao, NIU, Feiyu, GUO, Weibang, WU, Yilong
• Format: Article
• Language: Chinese
• Subjects: Deoxyribonucleic acid; DNA; Lung cancer; Mutation; Polymorphism
• ISSN: 1009-3419; 1999-6187
• DOI: 10.3779/cjlc.v14i10.3192

背景与目的 p53基因与人类多种肿瘤相关，突变型具有致癌作用，主要分布在外显子5-8。本研究 旨在建立高分辨熔解曲线（high resolution melting, HRM）检测非小细胞肺癌（non-small cell lung cancer, NSCLC）患者 p53基因突变的方法，探讨p53基因突变的特点及其在NSCLC发生发展中的演变规律。方法 采用HRM法检测264例 NSCLC患者肿瘤组织和54例癌旁肺组织p53基因外显子5-8的突变，突变样品进一步使用PCR产物直接测序法分析确 定突变类型；HRM法检测阳性而PCR产物直接测序法检测阴性的样品，进一步进行亚克隆测序证实。结果 54例对 照未发现突变。264例肿瘤组织中，HRM法检出 p53基因突变104例，102例得到PCR产物直接测序法证实，突变率为 39.4%；95例为点突变，7例为碱基插入和缺失导致的移码突变。p53外显子5-8的突变率分别为11.7%、8%、12.5%和 10.6%，差异无统计学意义（P=0.35）。p53基因突变与性别有关，与其它临床病理特征无关。结论 HRM法筛选p53 基因突变样品，具有操作简便、快速、敏感、单管避免污染等优点，值得推广。p53基因的突变特点提示，p53基因 突变是自发性突变，可能是DNA合成和修复过程中的随机错误所致。 Background and objective It has been proven that p53 gene was related to many human cancers. The mutations in p53 gene play an important role in carcinogensis and mostly happened in exon 5-8. The aim of this study is to establish a high resolution melting (HRM) assay to detect p53 mutations from patients with non-small cell lung cancer (NSCLC), to investigate the characteristics of p53 gene mutations, and to analyze the relationship between p53