Methylation Assessment for the Prediction of Malignancy in Mediastinal Adenopathies Obtained by Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration in Patients with Lung Cancer

Abstract

The evaluation of mediastinal lymph nodes is critical for the correct staging of patients with lung cancer (LC). Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a minimally invasive technique for mediastinal staging, though unfortunately lymph node micrometastasis is often missed by cytological analysis. The aim of this study was to evaluate the predictive capacity of methylation biomarkers and provide a classification rule for predicting malignancy in false negative EBUS-TBNA samples. The study included 112 patients with a new or suspected diagnosis of LC that were referred to EBUS-TBNA. Methylation of p16/INK4a, MGMT, SHOX2, E-cadherin, DLEC1, and RASSF1A was quantified by nested methylation-specific qPCR in 218 EBUS-TBNA lymph node samples. Cross-validated linear regression models were evaluated to predict malignancy. According to EBUS-TBNA and final diagnosis, 90 samples were true positives for malignancy, 110 were true negatives, and 18 were false negatives. MGMT, SHOX2, and E-cadherin were the methylation markers that better predicted malignancy. The model including sex, age, short axis diameter and standard uptake value of adenopathy, and SHOX2 showed 82.7% cross-validated sensitivity and 82.4% specificity for the detection of malignant lymphadenopathies among negative cytology samples. Our results suggest that the predictive model approach proposed can complement EBUS-TBNA for mediastinal staging.

Analysis of the Prognostic Value and Gene Expression Mechanism of SHOX2 in Lung Adenocarcinoma

“…et al, 2016). Additionally, there have been other similar reports of SHOX2 methylation as a predictor of malignancy (Leiro et al, 2019;Xu et al, 2020).…”

Comparison of Bisulfite Pyrosequencing and Methylation-Specific qPCR for Methylation Assessment

“…DNA quantity (DNAQ) using MYOD1 was obtained for each sample for normalization (standard curve 91.06% efficiency; slope: −3.5566, R 2 = 0.9985). Finally, the normalized methylation percentage (NMP) was calculated as follows [6]:…”

“…In lung cancer, hypermethylation of promoters of well-known cancer-related genes has been reported in many studies [2][3][4]. Aberrant methylation has been detected in liquid biopsy, including blood, sputum, bronchial washings, bronchoalveolar lavage, bronchial aspirates and cytohistological material from endobronchial ultrasound transbronchial needle aspiration (EBUS-TBNA) [4][5][6]. EBUS-TBNA is a minimally invasive method that allows the sampling of mediastinal and hilar lymph nodes for diagnosis and staging of lung cancer [7].…”

“…EBUS-TBNA is a minimally invasive method that allows the sampling of mediastinal and hilar lymph nodes for diagnosis and staging of lung cancer [7]. Samples obtained by EBUS-TBNA are typically analyzed using cytological techniques, though molecular markers, including epigenetics, are considered very promising for the detection of occult lymph node metastasis [6][7][8].…”

Detection and Independent Validation of Model-Based Quantitative Transcriptional Regulation Relationships Altered in Lung Cancers

“…Molecular biomarkers are usually evaluated for their individual associations with the disease risks and have been widely used in the decision processes of disease diagnosis and prognosis (He et al, 2019;Leiro et al, 2019;Yokobori et al, 2019;Muller et al, 2020). Transcription factor (TF) plays a critical role in cancer cell processes, such as cell proliferation, apoptosis, migration, and regulate gene expression.…”