Differential DNA Hypermethylation of Critical Genes Mediates the Stage-Specific Tobacco Smoke-Induced Neoplastic Progression of Lung Cancer

• Published in: Clinical cancer research 2005-04, Vol.11 (7), p.2466-2470
• Main Authors: Russo, Andrea L, Thiagalingam, Arunthathi, Pan, Hongjie, Califano, Joseph, Cheng, Kuang-hung, Ponte, Jose F, Chinnappan, Dharmaraj, Nemani, Pratima, Sidransky, David, Thiagalingam, Sam
• Format: Article
• Language: English
• Subjects: Apoptosis Regulatory Proteins; Biomarkers, Tumor - genetics; blood lymphocytes; Bronchi - metabolism; Bronchi - pathology; Cadherins - genetics; Calcium-Calmodulin-Dependent Protein Kinases - genetics; Carcinoma, Non-Small-Cell Lung - etiology; Carcinoma, Non-Small-Cell Lung - genetics; Cyclin-Dependent Kinase Inhibitor p16 - genetics; Death-Associated Protein Kinases; diagnostic markers; Disease Progression; DNA Methylation; DNA, Neoplasm - genetics; DNA, Neoplasm - metabolism; DNA-Binding Proteins - genetics; Gene Expression Regulation, Neoplastic; Glutathione S-Transferase pi; Glutathione Transferase - genetics; Humans; Isoenzymes - genetics; lung cancer; Lung Neoplasms - blood; Lung Neoplasms - etiology; Lung Neoplasms - genetics; Models, Biological; MSP; Neoplasm Staging; O-Methylguanine-DNA Methyltransferase - genetics; Polymerase Chain Reaction; Precancerous Conditions - genetics; Promoter Regions, Genetic - genetics; Respiratory Mucosa - metabolism; Respiratory Mucosa - pathology; Smad8 Protein; Smoking - adverse effects; tobacco smoke; Trans-Activators - genetics
• ISSN: 1078-0432; 1557-3265
• DOI: 10.1158/1078-0432.CCR-04-1962

Promoter DNA methylation status of six genes in samples derived from 27 bronchial epithelial cells and matching blood samples from 22 former/current smokers and five nonsmokers as well as 49 primary non–small cell lung cancer samples with corresponding blood controls was determined using methylation-specific PCR (MSP). Lung tumor tissues showed a significantly higher frequency of promoter DNA methylation in p16 , MGMT , and DAPK ( P < 0.05; Fisher's exact test). p16 promoter DNA methylation in tumors was observed at consistently higher levels when compared with all the other samples analyzed ( P = 0.001; Fisher's exact test). ECAD and DAPK exhibited statistically insignificant differences in their levels of DNA methylation among the tumors and bronchial epithelial cells from the smokers. Interestingly, similar levels of methylation were observed in bronchial epithelial cells and corresponding blood from smokers for all four genes ( ECAD , p16 , MGMT , and DAPK ) that showed smoking/lung cancer–associated methylation changes. In summary, our data suggest that targeted DNA methylation silencing of ECAD and DAPK occurs in the early stages and that of p16 and MGMT in the later stages of lung cancer progression. We also provide preliminary evidence that peripheral lymphocytes could potentially be used as a surrogate for bronchial epithelial cells to detect altered DNA methylation in smokers.