Detection of p53 gene mutations in exhaled breath condensate of non-small cell lung cancer patients
Detection of p53 gene mutations in exhaled breath condensate of non-small cell lung cancer patients
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DOI：
10.1016/j.lungcan.2003.08.034
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发表时间：
2004-02-01
期刊：
影响因子：
5.3
通讯作者：
Wirtz, H
中科院分区：
文献类型：
--
作者：
Gessner, C;Kuhn, H;Wirtz, H
肺癌的早期诊断是非常需要的。关于气道癌变过程的早期信息的一个潜在来源是呼出气冷凝物（EBC）。检测癌变的直接方法是检查慢性损伤区域（即气道和肺实质）的DNA。因此，我们研究了NSCLC患者和健康志愿者EBC中的DNA。通过PCR扩增呼出气冷凝液中的人DNA，并用于检测p53突变。β-肌动蛋白基因片段的PCR用于检测每个EBC样品中的人DNA。在65.7%的样本中，发现了β-肌动蛋白基因。提取的DNA以及天然EBC同样适合作为扩增的起始材料。在NSCLC患者的所有EBC样本中研究p53基因的突变。使用巢式PCR扩增p53外显子5-8并随后测序。在4名患者（n = 11; 36.4%）中发现突变，而在志愿者（n = 10）中未发现突变。EBC中检测到的突变也与相应的肿瘤组织进行了比较。所有病例均发现EBC和肿瘤组织中存在不同的点突变。我们的研究结果表明，呼出气冷凝物可用于分析直接烟草相关DNA损伤区域的体细胞基因突变。(C)2003爱思唯尔爱尔兰有限公司保留所有权利。
Early diagnosis of lung carcinoma is greatly desired. A potential source of early information regarding the process of cancerisation in the airways is exhaled breath condensate (EBC). The direct approach to detecting cancerisation is examining DNA from the area of chronic damage, i.e. airways and lung parenchyma. We therefore investigated DNA in EBC of patients with NSCLC and healthy volunteers. Human DNA was amplified by PCR in exhaled breath condensate and used to detect p53 mutations. A PCR of the beta-actin gene fragment was used to detect human DNA in each of the EBC samples. In 65.7% of the samples, the beta-actin gene was found. Extracted DNA as well as native EBC were equally suited as starting material for amplification. Mutations of the p53 gene were investigated in all EBC samples of NSCLC patients. p53 exons 5-8 were amplified using nested PCR and subsequently sequenced. Mutations were found in four of the patients (n = 11; 36.4%) while no mutation was found in volunteers (n = 10). Mutations detected in EBC were also compared with those of corresponding tumor tissue. Different point mutations in EBC and tumor tissue were revealed in all cases. Our findings demonstrate that exhaled breath condensate may be used for analysis of somatic gene mutations in an area of direct tobacco-related DNA damage. (C) 2003 Elsevier Ireland Ltd. All rights reserved.