title: Arecoline induced cell cycle arrest, apoptosis, and cytotoxicity to human endothelial cells.
abstract: Betel quid (BQ) chewing is a common oral habit in South Asia and Taiwan. BQ consumption may increase the risk of oral squamous cell carcinoma (OSCC), oral submucous fibrosis (OSF), and periodontitis as well as systemic diseases (atherosclerosis, hypertension, etc.). However, little is known about the toxic effect of BQ components on endothelial cells that play important roles for angiogenesis, carcinogenesis, tissue fibrosis, and cardiovascular diseases. EAhy 926 (EAHY) endothelial cells were exposed to arecoline, a major BQ alkaloid, for various time periods. Cytotoxicity was estimated by 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. The cell cycle distribution of EAHY cells residing in sub-G0/G1, G0/G1, S-, and G2/M phases was analyzed by propidium iodide staining of cellular DNA content and flow cytometry. Some EAHY cells retracted, became round-shaped in appearance, and even detached from the culture plate after exposure to higher concentrations of arecoline (> 0.4 mM). At concentrations of 0.4 and 0.8 mM, arecoline induced significant cytotoxicity to EAHY cells. At similar concentrations, arecoline induced G2/M cell cycle arrest and increased sub-G0/G1 population, a hallmark of apoptosis. Interestingly, prolonged exposure to arecoline (0.1 mM) for 12 and 21 days significantly suppressed the proliferation of EAHY cells, whereas EAHY cells showed adaptation and survived when exposed to 0.05 mM arecoline. These results suggest that BQ components may contribute to the pathogenesis of OSF and BQ chewing-related cardiovascular diseases via toxicity to oral or systemic endothelial cells, leading to impairment of vascular function. During BQ chewing, endothelial damage may be induced by areca nut components and associate with the pathogenesis of OSF, periodontitis, and cardiovascular diseases.
authors.affiliation: Graduate Institute of Clinical Dentistry, National Yang-Ming University, No. 155, Sec. 2, Li-Nong St, Beitou District, Taipei, Taiwan.
authors.forename: Shuei-Kuen
authors.initials: SK
authors.lastname: Tseng
authors.forename: Mei-Chi
authors.initials: MC
authors.lastname: Chang
authors.forename: Cheng-Yao
authors.initials: CY
authors.lastname: Su
authors.forename: Lin-Yang
authors.initials: LY
authors.lastname: Chi
authors.forename: Jenny Zwei-Ching
authors.initials: JZ
authors.lastname: Chang
authors.forename: Wan-Yu
authors.initials: WY
authors.lastname: Tseng
authors.forename: Sin-Yuet
authors.initials: SY
authors.lastname: Yeung
authors.forename: Ming-Lun
authors.initials: ML
authors.lastname: Hsu
authors.forename: Jiiang-Huei
authors.initials: JH
authors.lastname: Jeng
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001115:Arecoline / Q000633:toxicity*; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018679:Cholinergic Agonists / Q000633:toxicity*; D004396:Coloring Agents; D004247:DNA / Q000032:analysis; D042783:Endothelial Cells / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000166:cytology; D005434:Flow Cytometry; D016193:G1 Phase / Q000187:drug effects; D006801:Humans; D007399:Interphase / Q000187:drug effects; D011419:Propidium; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors
chemical_list: D018679:Cholinergic Agonists; D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D011419:Propidium; D001115:Arecoline; D004247:DNA; C022616:thiazolyl blue