title: Cigarette smoke extract induces aberrant cytochrome-c oxidase subunit II methylation and apoptosis in human umbilical vascular endothelial cells.
abstract: Cigarette smoke-induced apoptosis of vascular endothelial cells contributes to the pathogenesis of chronic obstructive pulmonary disease. However, the mechanisms responsible for endothelial apoptosis remain poorly understood. We conducted an in vitro study to investigate whether DNA methylation is involved in smoking-induced endothelial apoptosis. Human umbilical vascular endothelial cells (HUVECs) were exposed to cigarette smoke extract (CSE) at a range of concentrations (0-10%). HUVECs were also incubated with a demethylating reagent, 5-aza-2'-deoxycytidinem (AZA), with and without CSE. Apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay and flow cytometry using annexin V-FITC/propidium iodide staining. We found that CSE treatment significantly increased HUVEC apoptosis in a dose- and time-dependent manner. Quantitative real-time RT-PCR and immunoblot revealed that CSE treatment decreased cytochrome-c oxidase subunit II (COX II) mRNA and protein levels and decreased COX activity. Methylation-specific PCR and direct bisulfite sequencing revealed positive COX II gene methylation. AZA administration partly increased mRNA and protein expressions of COX II, and COX activity decreased by CSE and attenuated the toxic effects of CSE. Our results showed that CSE induced aberrant COX II methylation and apoptosis in HUVECs.
authors.affiliation: Department of Respiratory Medicine, The Second Xiangya Hospital of Central-South University, Hunan, Changsha, China;
authors.forename: Min
authors.initials: M
authors.lastname: Yang
authors.affiliation: Department of Respiratory Medicine, The Second Xiangya Hospital of Central-South University, Hunan, Changsha, China; pingchen0731@sina.com.
authors.forename: Ping
authors.initials: P
authors.lastname: Chen
authors.affiliation: Department of Respiratory Medicine, The Second Xiangya Hospital of Central-South University, Hunan, Changsha, China;
authors.forename: Hong
authors.initials: H
authors.lastname: Peng
authors.affiliation: Emergency Department, The Second Xiangya Hospital of Central-South University, Hunan, Changsha, China;
authors.forename: Hongliang
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Respiratory Medicine, The Second Xiangya Hospital of Central-South University, Hunan, Changsha, China;
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Respiratory Medicine, The Second Xiangya Hospital of Central-South University, Hunan, Changsha, China;
authors.forename: Shan
authors.initials: S
authors.lastname: Cai
authors.affiliation: Dermatological Department, The Second Xiangya Hospital of Central-South University, Hunan, Changsha, China;
authors.forename: Qianjin
authors.initials: Q
authors.lastname: Lu
authors.affiliation: Department of Physiology, Central-South University, Hunan, Changsha, China.
authors.forename: Chaxiang
authors.initials: C
authors.lastname: Guan
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001483:Base Sequence; D002478:Cells, Cultured; D003576:Electron Transport Complex IV / Q000378:metabolism*; D004790:Enzyme Induction / Q000187:drug effects / Q000502:physiology; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D008745:Methylation / Q000187:drug effects; D008969:Molecular Sequence Data; D012906:Smoke / Q000009:adverse effects*; D014026:Nicotiana / Q000633:toxicity*
chemical_list: D012906:Smoke; C117108:cytochrome C oxidase subunit II; D003576:Electron Transport Complex IV
keywords: DNA methylation; apoptosis; cigarette smoke; cytochrome-c oxidase subunit II; endothelial cells