中国肺癌杂志2011，Vol.14Issue(10)：767-773,7.DOI:10.3779/j.issn.1009-3419.2011.10.01
高分辨熔解曲线法检测非小细胞肺癌p53基因的突变
High Resolution Melting Analysis for Detecting p53 Gene Mutations in Patients with Non-small Cell Lung Cancer
陈志红 1吴一龙 1安社娟 1谢至 1严红虹 1陈剑光 1苏健 1张绪超 1牛飞玉 1郭伟浜1
作者信息
- 1. 510080,广州,广东省人民医院医学研究中心,广东省医学科学院,广东省肺癌研究所
- 折叠
摘要
Abstract
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 es-tablish 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 mutations and evolution regularity of pathogenesis. Methods p53 mutations in exon 5-8 were detected by HRM assay on DNA insolated from 264 NSCLC samples derived from tumor tissues and 54 control samples from pericancerous pulmonary tissues. The mutation samples by the HRM assay were confirmed by sequencing technique. Samples which were positive by HRM but wild type by sequencing were further confirmed by sub-clone and sequencing. Results No mutation was found in 54 pericancerous pul-monary samples by HRM assay. 104 of the 264 tumor tissues demonstrated mutation curves by HRM assay, 102 samples were confirmed by sequencing, including 95 point mutations and 7 frame shift mutations by insertion or deletion. The mutation rate of p53 gene was 39.4%. The mutation rate from exon 5-8 were 11.7%, 8%, 12.5% and 10.6%, respectively and there was no statistically significant difference between them (P=0.35). p53 mutations were significantly more frequent in males than that in females, but not related to the other clinicopathologic characteristics. Conclusion The results indicate that HRM is a sensitive in-tube methodology to detect for mutations in clinical samples. The results suggest that the arising p53 mutations in NSCLC may be due to spontaneous error in DNA synthesis and repair.关键词
p53基因/突变/测序Key words
p53 gene/ Mutation/ Sequence分类
医药卫生引用本文复制引用
陈志红,吴一龙,安社娟,谢至,严红虹,陈剑光,苏健,张绪超,牛飞玉,郭伟浜..高分辨熔解曲线法检测非小细胞肺癌p53基因的突变[J].中国肺癌杂志,2011,14(10):767-773,7.
