Chang M.-C.LI-DEH LINWu H.-L.Ho Y.-S.Hsien H.-C.TONG-MEI WANGJeng P.-Y.Cheng R.-H.Hahn L.-J.JIIANG-HUEI JENG2021-07-052021-07-0520130143-3334https://www.scopus.com/inward/record.uri?eid=2-s2.0-84877592734&doi=10.1093%2fcarcin%2fbgt012&partnerID=40&md5=315055634bd354146ab9d5a64f5ff545https://scholars.lib.ntu.edu.tw/handle/123456789/569280Betel quid (BQ) chewing is an oral habit that increases the risk of oral cancer and oral submucous fibrosis (OSF), a precancerous condition showing epithelial atrophy and tissue fibrosis. Persistent fibroblast contraction may induce the fibrotic contracture of tissue. In this study, we found that areca nut extract (ANE) (200-1200 μg/ ml) stimulated buccal mucosa fibroblast (OMF)-populated collagen gel contraction. Arecoline but not arecaidine-two areca alkaloids, slightly induced the OMF contraction. Exogenous addition of carboxylesterase (2 U/ml) prevented the arecoline- but not ANEinduced OMF contraction. OMF expressed inositol triphosphate (IP3) receptors. ANE-induced OMF (800 μg/ml) contraction was inhibited by U73122 [phospholipase C (PLC) inhibitor] and 2-aminoethoxydiphenyl borate (IP3 receptor antagonist), respectively. Ethylene glycol tetraacetic acid and verapamil, two calcium mobilization modulators, also suppressed the ANE-induced OMF contraction. ANE induced calcium/calmodulin kinase II and myosin light chain (MLC) phosphorylation in OMF. Moreover, W7 (a Ca2+/ calmodulin inhibitor), HA1077 (Rho kinase inhibitor), ML-7 (MLC kinase inhibitor) and cytochalasin B (actin filament polymerization inhibitor) inhibited the ANE-induced OMF contraction. Although ANE elevated reactive oxygen species (ROS) level in OMF, catalase, superoxide dismutase and N-acetyl-l-cysteine showed no obvious effect on ANE-elicited OMF contraction. These results indicate that BQ chewing may affect the wound healing and fibrotic processes in OSF via inducing OMF contraction by ANE and areca alkaloids. AN components-induced OMF contraction was related to PLC/ IP3/Ca2+/calmodulin and Rho signaling pathway as well as actin filament polymerization, but not solely due to ROS production. ? The Author 2013. Published by Oxford University Press. All rights reserved.[SDGs]SDG3acetylcysteine; arecaidine; arecoline; calcium calmodulin dependent protein kinase II; calcium ion; calmodulin; carboxylesterase; catalase; collagen gel; cytochalasin B; egtazic acid; inositol trisphosphate; messenger RNA; myosin light chain; reactive oxygen metabolite; Rho kinase; superoxide dismutase; verapamil; article; betel nut; cheek mucosa; controlled study; enzyme activation; enzyme phosphorylation; fibroblast; human; human tissue; mouth disease; polymerization; priority journal; protein expression; wound healing; Areca; Arecoline; Calcium; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Cells, Cultured; Fibroblasts; Humans; Inositol 1,4,5-Trisphosphate Receptors; Mouth Mucosa; Mouth Neoplasms; Myosin Light Chains; Nuts; Oral Submucous Fibrosis; Phosphorylation; Plant Extracts; Precancerous Conditions; Reactive Oxygen Species; Signal Transductionjournal article10.1093/carcin/bgt012233490212-s2.0-84877592734