Oh Tae Jeong, Jang Seunghyun, Kim Su Ji, Woo Min A, Son Ji Woong, Jeong In Beom, Lee Min Hyeok, An Sungwhan
Genomictree, Inc., Daejeon 34027, Republic of Korea.
Department of Internal Medicine, Konyang University Hospital, Daejeon 35365, Republic of Korea.
Oncol Lett. 2024 Apr 3;27(6):246. doi: 10.3892/ol.2024.14379. eCollection 2024 Jun.
Bronchoscopy is a frequently used initial diagnostic procedure for patients with suspected lung cancer (LC). Cytological examinations of bronchial washing (BW) samples obtained during bronchoscopy often yield inconclusive results regarding LC diagnosis. The present study aimed to identify molecular biomarkers as a non-invasive method for LC diagnosis. Aberrant DNA methylation is used as a useful biomarker for LC. Therefore, microarray-based methylation profiling analyses on 13 patient-matched tumor tissues at stages I-III vs. non-tumor tissues were performed, and a group of highly differentially methylated genes was identified. A subsequent analysis using bisulfite-pyrosequencing with additional tissues and cell lines revealed six methylated genes [ADAM metallopeptidase with thrombospondin type 1 motif 20, forkhead box C2 (mesenchyme forkhead 1), NK2 transcription factor related, locus 5 (), oligodendrocyte transcription factor 3, protocadherin γ subfamily A 12 () and paired related homeobox 1 ()] associated with LC. Next, a highly sensitive and accurate detection method, linear target enrichment-quantitative methylation-specific PCR in a single closed tube, was applied for clinical validation using BW samples from patients with LC (n=68) and individuals with benign diseases (n=33). and methylation were identified as the best-performing biomarkers to detect LC. The two-marker combination showed a sensitivity of 82.4% and a specificity of 87.9%, with an area under the curve of 0.891. Notably, the sensitivity for small cell LC was 100%. The two-marker combination had a positive predictive value of 93.3% and a negative predictive value of 70.7%. The sensitivity was higher than that of cytology, which only had a sensitivity of 50%. The methylation status of the two-marker combination showed no association with sex, age or stage, but was associated with tumor location and histology. In conclusion, the present study showed that the regulatory regions of and are highly methylated in LC and can be used to detect LC in BW specimens as a diagnostic adjunct to cytology in clinical practice.
支气管镜检查是疑似肺癌（LC）患者常用的初始诊断方法。支气管镜检查期间获取的支气管冲洗（BW）样本的细胞学检查在LC诊断方面往往结果不明确。本研究旨在确定分子生物标志物作为LC诊断的非侵入性方法。异常DNA甲基化被用作LC的有用生物标志物。因此，对13例I - III期患者匹配的肿瘤组织与非肿瘤组织进行了基于微阵列的甲基化谱分析，并鉴定出一组高度差异甲基化基因。随后使用亚硫酸氢盐焦磷酸测序对额外的组织和细胞系进行分析，发现了六个与LC相关的甲基化基因[含血小板反应蛋白基序的金属蛋白酶20、叉头框C2（间充质叉头1）、NK2转录因子相关位点5、少突胶质细胞转录因子3、原钙黏蛋白γ亚家族A12和配对相关同源盒1]。接下来，一种高灵敏度和准确性的检测方法，即单管线性靶标富集 - 定量甲基化特异性PCR，被应用于使用LC患者（n = 68）和良性疾病个体（n = 33）的BW样本进行临床验证。 和 甲基化被确定为检测LC的最佳生物标志物。双标志物组合的灵敏度为82.4%，特异性为87.9%，曲线下面积为0.891。值得注意的是，对小细胞LC的灵敏度为100%。双标志物组合的阳性预测值为93.3%，阴性预测值为70.7%。其灵敏度高于细胞学检查，后者的灵敏度仅为50%。双标志物组合的甲基化状态与性别、年龄或分期无关，但与肿瘤位置和组织学有关。总之，本研究表明， 在LC中高度甲基化，可用于在BW标本中检测LC，作为临床实践中细胞学诊断的辅助手段。
