Wang Tong-Hong, Shen Yen-Wen, Chen Hsin-Ying, Chen Chih-Chieh, Lin Nan-Chin, Shih Yin-Hwa, Hsia Shih-Min, Chiu Kuo-Chou, Shieh Tzong-Ming
Biobank, Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan.
School of Dentistry, China Medical University, Taichung 404328, Taiwan.
Biomedicines. 2024 Feb 9;12(2):412. doi: 10.3390/biomedicines12020412.
Areca nut is a major contributor to the high prevalence of oral cancer in Asia. The precise mechanisms by which areca nut stimulates mucosal cells and contributes to the progression of oral cancer urgently require clarification. The current study aimed to assess the effects of arecoline on the normal human gingival epithelium cell line S-G. Cell viability, levels of reactive oxygen species (ROS), protein expression, cellular morphology, and gene expression were evaluated using the MTT test, flow cytometry, Western blot analysis, optical or confocal microscopy, and RT-qPCR. Keratin (KRT6) analysis involved matched normal and cancer tissues from clinical head and neck specimens. The results demonstrated that 12.5 µg/mL of arecoline induced ROS production, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) mRNA expression in S-G cells. This activation of the MAPK/ERK pathway increased KRT6 expression while limiting cell migration. In head and neck cancer tissues, gene expression exceeded that of normal tissues. This study confirms that arecoline induces ROS accumulation in normal cells, leading to the secretion of proinflammatory factors and KRT6 expression. This impedes oral mucosal healing, thereby promoting the progression of oral cancer.
槟榔是亚洲口腔癌高发病率的主要促成因素。槟榔刺激黏膜细胞并促进口腔癌进展的确切机制迫切需要阐明。当前研究旨在评估槟榔碱对正常人牙龈上皮细胞系S-G的影响。使用MTT试验、流式细胞术、蛋白质印迹分析、光学或共聚焦显微镜以及RT-qPCR评估细胞活力、活性氧（ROS）水平、蛋白质表达、细胞形态和基因表达。角蛋白（KRT6）分析涉及来自临床头颈标本的配对正常组织和癌组织。结果表明，12.5 µg/mL的槟榔碱诱导S-G细胞中ROS生成、肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）mRNA表达。MAPK/ERK通路的这种激活增加了KRT6表达，同时限制了细胞迁移。在头颈癌组织中，基因表达超过正常组织。本研究证实槟榔碱诱导正常细胞中ROS积累，导致促炎因子分泌和KRT6表达。这会阻碍口腔黏膜愈合，从而促进口腔癌进展。
