Ras联合域家族1A亚型（RASSF1A）和矮人家族转录因子2（SHOX2）的异常甲基化已被验证为诊断早期肺腺癌（LUADs）的一对有价值的生物标志物。表皮生长因子受体（EGFR）是肺癌发生的关键驱动突变。本研究旨在调查258个早期LUADs标本中RASSF1A和SHOX2的异常启动子甲基化和EGFR的遗传突变。我们回顾性地选择了258个直径小于2厘米的肺结节的石蜡包埋样品，并评估了单个生物标志物检测和非侵入性（第1组）和侵入性病变（第2A和2B组）之间的多个面板的诊断性能。然后，我们调查了遗传和表观遗传变化之间的相互作用。RASSF1A和SHOX2启动子甲基化以及EGFR突变的程度在侵袭性病变中明显高于非侵入性病变。三种生物标志物可可靠地区分非侵入性和侵入性病变，具有60.9％的灵敏度[95％信​​心区间（CI）为52.41–68.78％]和80.0％的特异性（95％ CI 为72.14–86.07％）。新的面板生物标志物可以进一步区分三种侵袭性的病理亚型（曲线下面积值> 0.6）。RASSF1A甲基化和EGFR突变的分布在早期LUAD中是相当独特的（P = 0.002）。RASSF1A和SHOX2的DNA甲基化是一对有前途的生物标志物，可以与EGFR突变等其他驱动变化结合使用，以支持LUADs的不同诊断，尤其是I期。©2023年。本文作者授予Springer-Verlag GmbH Germany独家许可，属于Springer Nature的一部分。

Aberrant methylation of Ras association domain family 1, isoform A (RASSF1A), and short-stature homeobox gene 2 (SHOX2) promoters has been validated as a pair of valuable biomarkers for diagnosing early lung adenocarcinomas (LUADs). Epidermal growth factor receptor (EGFR) is the key driver mutation in lung carcinogenesis. This study aimed to investigate the aberrant promoter methylation of RASSF1A and SHOX2, and the genetic mutation of EGFR in 258 specimens of early LUADs.We retrospectively selected 258 paraffin-embedded samples of pulmonary nodules measuring 2 cm or less in diameter and evaluated the diagnostic performance of individual biomarker assays and multiple panels between noninvasive (group 1) and invasive lesions (groups 2A and 2B). Then, we investigated the interaction between genetic and epigenetic alterations.The degree of RASSF1A and SHOX2 promoter methylation and EGFR mutation was significantly higher in invasive lesions than in noninvasive lesions. The three biomarkers distinguished between noninvasive and invasive lesions with reliable sensitivity and specificity: 60.9% sensitivity [95% confidence interval (CI) 52.41-68.78] and 80.0% specificity (95% CI 72.14-86.07). The novel panel biomarkers could further discriminate among three invasive pathological subtypes (area under the curve value > 0.6). The distribution of RASSF1A methylation and EGFR mutation was considerably exclusive in early LUAD (P = 0.002).DNA methylation of RASSF1A and SHOX2 is a pair of promising biomarkers, which may be used in combination with other driver alterations, such as EGFR mutation, to support the differential diagnosis of LUADs, especially for stage I.© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.