Zhou Caicun, Ni Jian, Zhao Yinmin, Su Bo
Department of Oncology, Shanghai Pulmonary Hospital, Tongji University, Shanghai, China.
Cancer J. 2006 Jan-Feb;12(1):33-9. doi: 10.1097/00130404-200601000-00007.
We investigated somatic mutations of the epidermal growth factor receptor gene in non-small cell lung cancer tumor tissue and their detection using real-time polymerase chain reaction with TaqMan-MGB probes.
The DNA was extracted from surgically resected non-small cell lung cancer tumor specimens. Genes encoding for epidermal growth factor receptor tyrosine (exons 18, 19, and 21) were amplified by nested polymerase chain reaction, sequenced, and analyzed by chromatograms with manual review. TaqMan-MGB probes were designed to detect the epidermal growth factor receptor gene mutations in the tumor tissues using real-time polymerase chain reaction.
Somatic point mutations and deletions were identified in the tyrosine kinase domain of the epidermal growth factor receptor gene in 21 of 80 non-small cell lung cancer patients, including 13 patients with deletion mutations occurring in exon 19 and 8 patients with point mutations occurring in codon 858 (exon 21). The results from real-time polymerase chain reaction with TaqMan-MGB probes were completely consistent with sequencing outcomes. Both the sensitivity and specificity for detecting the epidermal growth factor receptor gene mutations using real-time polymerase chain reaction with TaqMan-MGB probes were 100%. The mutation incidence was significantly higher in female patients, nonsmokers, and patients with adenocarcinoma than in male patients, smokers, and those with nonadenocarcinomas (P < 0.05). The mutations were not related to patient's age or tumor nodal metastasis staging.
Somatic mutations of the epidermal growth factor receptor gene that develop in non-small cell lung cancer patients are more common in female patients, nonsmokers, and patients with adenocarcinoma. Real-time polymerase chain reaction using TaqMan-MGB probes is effective, simple, and fast in the detection of epidermal growth factor receptor gene mutations.
我们研究了非小细胞肺癌肿瘤组织中表皮生长因子受体基因的体细胞突变，以及使用TaqMan-MGB探针通过实时聚合酶链反应对其进行检测的情况。
从手术切除的非小细胞肺癌肿瘤标本中提取DNA。通过巢式聚合酶链反应扩增编码表皮生长因子受体酪氨酸（外显子18、19和21）的基因，进行测序，并通过色谱图进行分析且人工复核。设计TaqMan-MGB探针，使用实时聚合酶链反应检测肿瘤组织中的表皮生长因子受体基因突变。
在80例非小细胞肺癌患者中的21例患者的表皮生长因子受体基因酪氨酸激酶结构域中鉴定出体细胞点突变和缺失，其中13例患者在外显子19中发生缺失突变，8例患者在密码子858（外显子21）中发生点突变。使用TaqMan-MGB探针进行实时聚合酶链反应的结果与测序结果完全一致。使用TaqMan-MGB探针通过实时聚合酶链反应检测表皮生长因子受体基因突变的敏感性和特异性均为100%。女性患者、不吸烟者和腺癌患者的突变发生率显著高于男性患者、吸烟者和非腺癌患者（P<0.05）。这些突变与患者年龄或肿瘤淋巴结转移分期无关。
非小细胞肺癌患者中发生的表皮生长因子受体基因体细胞突变在女性患者、不吸烟者和腺癌患者中更为常见。使用TaqMan-MGB探针的实时聚合酶链反应在检测表皮生长因子受体基因突变方面有效、简便且快速。
