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), p.767-773
• 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: Adult; Aged; Aged, 80 and over; Carcinoma, Non-Small-Cell Lung - genetics; Chi-Square Distribution; DNA Mutational Analysis - methods; DNA, Neoplasm - chemistry; DNA, Neoplasm - genetics; Female; Humans; Lung Neoplasms - genetics; Male; Middle Aged; Mutation; Mutation Rate; Nucleic Acid Denaturation; p53 gene; Polymerase Chain Reaction - methods; Sequence; Transition Temperature; Tumor Suppressor Protein p53 - genetics; Young Adult
• ISSN: 1009-3419; 1999-6187
• DOI: 10.3779/j.issn.1009-3419.2011.10.01

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 mutations and evolution regularity of pathogenesis. 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. No mutation was found in 54 pericancerous pulmonary samples by HRM assay. 104 of the 264 tumor tissues demonstrated mutation curves by HRM assay, 102 samples were confirmed b