Department of Chest Surgery, Oncologic and General Surgery, University Hospital No. 2, Zeromskiego St. 113, 91-647, Medical University of Lodz, Poland.
Respir Med. 2012 Sep;106(9):1293-300. doi: 10.1016/j.rmed.2012.06.019. Epub 2012 Jul 13.
Exhaled breath condensate (EBC) contains extracellular DNA that may originate from pathological lesions of the respiratory tract and can be a genetic marker of pulmonary malignancy. We tested whether complete surgical excision of lung cancer will decrease exhalation of mutated KRAS oncogene. Fifty seven patients with clinical diagnosis of lung cancer and detectable KRAS mutations in pre-surgery EBC-DNA were qualified for surgical treatment. Point mutations at codon 12 of KRAS oncogene were detected using mutant-enriched PCR technique in DNA from pre-surgery blood, EBC collected before, 7 and 30 days after surgery and from specimens of resected tumor and normal pulmonary parenchyma. The ratio of mutated to wild type KRAS DNA (R mut/wild KRAS) was calculated for each specimen after electrophoresis and densitometry of the final amplification and digestion product. In 46 patients non-small cell lung cancer (NSCLC) and in 11 benign lesion (BL) were confirmed. All blood and tumor specimens were positive for KRAS mutations, while 41 specimens of normal pulmonary parenchyma were negative. In NSCLC patients pre-surgery EBC R mut/wild KRAS of 0.20 ± 0.03 decreased by 1.3- and 3.7-times (p < 0.001) at 7th and 30th day and 10 EBC specimens at day 30th became negative. The highest R mut/wild KRAS was found in NSCLC specimens - 1.36 ± 0.29 while the lowest in pulmonary parenchyma - 0.02 ± 0.03 (p < 0.001). R mut/wild KRAS in EBC did not correlate with the blood and cancer ratios. Determination of mutated KRAS oncogene in EBC can be potentially helpful in the follow-up of surgical treatment of pulmonary malignancy.
呼出气冷凝物(EBC)含有可能源自呼吸道病变的细胞外 DNA,并且可以作为肺恶性肿瘤的遗传标志物。我们检测了肺癌的完全手术切除是否会减少呼出的突变 KRAS 致癌基因。57 名临床诊断为肺癌且术前 EBC-DNA 中可检测到 KRAS 突变的患者有资格接受手术治疗。使用突变富集 PCR 技术在术前血液、手术前、手术后 7 天和 30 天采集的 EBC 以及切除的肿瘤和正常肺组织标本的 DNA 中检测 KRAS 致癌基因 12 号密码子的点突变。在电泳和最终扩增和消化产物的密度测量后,为每个标本计算突变型与野生型 KRAS DNA 的比值(R mut / wild KRAS)。在 46 名非小细胞肺癌(NSCLC)患者和 11 名良性病变(BL)患者中得到证实。所有血液和肿瘤标本均为 KRAS 突变阳性,而 41 份正常肺组织标本均为阴性。在 NSCLC 患者中,术前 EBC 的 R mut / wild KRAS 为 0.20 ± 0.03,在第 7 天和第 30 天降低了 1.3- 和 3.7 倍(p <0.001),第 30 天的 10 个 EBC 标本变为阴性。在 NSCLC 标本中发现的 R mut / wild KRAS 最高,为 1.36 ± 0.29,而在肺组织中发现的最低,为 0.02 ± 0.03(p <0.001)。EBC 中的 R mut / wild KRAS 与血液和癌症比率无关。EBC 中突变 KRAS 致癌基因的测定可能有助于肺恶性肿瘤手术治疗的随访。