title: Chronic exposure to cigarette smoke condensate in vitro induces epithelial to mesenchymal transition-like changes in human bronchial epithelial cells, BEAS-2B.
abstract: Cigarette smoke causes lung tumorigenesis; however, the mechanisms underlying transformation are unknown. We investigated if tobacco compounds induce DNA promoter hypermethylation in BEAS-2B cells treated with low doses of cigarette smoke condensate (CSC) for one month. Transcriptional profiles and anchorage-independent growth were explored using Affymetrix microarray and soft agar assay, respectively. To investigate if tobacco compounds induce hypermethylation, CSC/dimethyl sulfoxide (DMSO)-treated cells were further treated with 5-Aza-2'-deoxycytidine (5AzaC) and trychostatin A (TSA). This treatment was followed by transcriptional profiling. CSC-exposed cells acquired a fibroblast-like shape with enhanced anchorage-independent growth. Silencing of epithelial cadherin, the hallmark of epithelial to mesenchymal transition (EMT), was observed upon exposure to CSC. Changes in the expression of genes involved in epidermal development, intercellular junction formation, and cytoskeleton formation were identified. Gene expression profiles from 5AzaC- and TSA-treated cells revealed 130 genes possibly methylated due to chronic CSC exposure. Our results suggest that E-cadherin may also be silenced by hypermethylation in an in vitro model of chronic exposure to low doses of CSC. This study demonstrates evidence for a tobacco compound induced EMT-like process in vitro and provides insight into possible mechanisms of gene silencing occurring during this treatment.
authors.affiliation: Philip Morris International R&D, Philip Morris Products SA, Neuchâtel, Switzerland. emilija.veljkovic@pmintl.com
authors.forename: Emilija
authors.initials: E
authors.lastname: Veljkovic
authors.forename: Josef
authors.initials: J
authors.lastname: Jiricny
authors.forename: Mirco
authors.initials: M
authors.lastname: Menigatti
authors.forename: Hubert
authors.initials: H
authors.lastname: Rehrauer
authors.forename: Wanjiang
authors.initials: W
authors.lastname: Han
mesh_terms: D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D001980:Bronchi / Q000473:pathology*; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000502:physiology; D019175:DNA Methylation; D000077209:Decitabine; D004847:Epithelial Cells / Q000473:pathology*; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D020869:Gene Expression Profiling; D005960:Glucosides / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007365:Intercellular Junctions / Q000502:physiology; D012906:Smoke / Q000009:adverse effects*
chemical_list: D005960:Glucosides; D006877:Hydroxamic Acids; D012906:Smoke; D000077209:Decitabine; C053043:trichostatin C; D001374:Azacitidine