Elevated vimentin expression in buccal mucosal fibroblasts by arecoline in vitro as a possible pathogenesis for oral submucous fibrosis.
Areca quid chewing is strongly correlated with oral submucous fibrosis (OSF) in Taiwan. The cytotoxicity of arecoline, a major areca nut alkaloid, on human oral fibroblasts has been extensively studied. To date, however, there has been little research exploring the possible effects of arecoline on cytoskeleton components. In this study, in addition to conducting a cytotoxicity assay, we examine the effect of arecoline on vimentin, an intermediate filament, and its expression in human buccal mucosal fibroblasts on exposure to various levels of arecoline (0-200 microg/ml) for 48 h. At a concentration above 50 microg/ml, arecoline demonstrated dose-dependent cytotoxicity (P<0.05) for cultured fibroblasts. Using sodium dodecyl sulphate-polyacrylamide gel electrophoresis, we demonstrated dose-dependent elevation of 57 kDa cytoskeletal-protein levels for arecoline. Evidence from immunoblotting assay indicated this 57 kDa cytoskeletal protein was vimentin. The increase in vimentin with arecoline exposure corresponded to that noted for fibroblasts cultured from OSF patients. Immunohistochemical assay also revealed that vimentin expression was much higher for OSF specimens than for normal buccal mucosa. We suggest these results may advance understanding of the possible pathogenesis for submucous fibrosis through the transformation of normal buccal mucosa as a result of areca quid chewing.
Department of Dentistry, Chung Shan Medical and Dental College Hospital, Taichung, Taiwan.
Yu-Chao
YC
Chang
Chung-Hung
CH
Tsai
Kuo-Wei
KW
Tai
Shyh-Hwang
SH
Yang
Ming-Yung
MY
Chou
Chong-Kuei
CK
Lii
D001115:Arecoline / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002610:Cheek; D018679:Cholinergic Agonists / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005260:Female; D005347:Fibroblasts / Q000737:chemistry / Q000187:drug effects*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D009061:Mouth Mucosa / Q000737:chemistry / Q000187:drug effects*; D009914:Oral Submucous Fibrosis / Q000139:chemically induced* / Q000378:metabolism; D014746:Vimentin / Q000032:analysis*
D018679:Cholinergic Agonists; D014746:Vimentin; D001115:Arecoline; D007770:L-Lactate Dehydrogenase