Australian Centre for Oral Oncology Research & Education, UWA Dental School, University of Western Australia, Nedlands, WA, Australia.
University of Queensland Centre for Clinical Research, The University of Queensland, Herston, QLD, Australia.
J Oral Pathol Med. 2018 May;47(5):511-518. doi: 10.1111/jop.12704. Epub 2018 Mar 22.
The role of alcohol-containing mouthwash as a risk factor for the development of oral cancer is a subject of conflicting epidemiological evidence in the literature despite alcohol being a recognised carcinogen. The aim of this study was to use in vitro models to investigate mechanistic and global gene expression effects of exposure to alcohol-containing mouthwash.
Two brands of alcohol-containing mouthwash and their alcohol-free counterparts were used to treat two oral cell lines derived from normal (OKF6-TERT) and dysplastic (DOK) tissues. Genotoxicity was determined by Comet assay. RNA-seq was performed using the Ion Torrent platform. Bioinformatics analysis used R/Bioconductor packages with differential expression using DEseq2. Pathway enrichment analysis used EnrichR with the WikiPathways and Kegg databases.
Both cell lines displayed dose-dependent DNA damage in response to acute exposure to ethanol and alcohol-containing mouthwashes as well as alcohol-free mouthwashes reconstituted with ethanol as shown by Comet assay. The transcriptomic effects of alcohol-containing mouthwash exposure were more complex with significant differential gene expression ranging from >2000 genes in dysplastic (DOK) cells to <100 genes in normal (OKF6-TERT) cells. Pathway enrichment analysis in DOK cells revealed alcohol-containing mouthwashes showed common features between the two brands used including DNA damage response as well as cancer-associated pathways. In OKF6-TERT cells, the most significantly enriched pathways involved inflammatory signalling.
Alcohol-containing mouthwashes are genotoxic in vitro to normal and dysplastic oral keratinocytes and induce widespread changes in gene expression. Dysplastic cells are more susceptible to the transcriptomic effects of mouthwash.
尽管酒精是一种公认的致癌物质，但含酒精漱口水作为口腔癌发展的危险因素，其在文献中的流行病学证据存在冲突。本研究旨在使用体外模型来研究暴露于含酒精漱口水后的机制和全基因表达效应。
使用两种品牌的含酒精漱口水及其无酒精对应物处理两种源自正常（OKF6-TERT）和发育不良（DOK）组织的口腔细胞系。通过彗星试验测定遗传毒性。使用 Ion Torrent 平台进行 RNA-seq。使用 R/Bioconductor 包进行生物信息学分析，使用 DEseq2 进行差异表达分析。通路富集分析使用 EnrichR 与 WikiPathways 和 Kegg 数据库。
两种细胞系在急性暴露于乙醇和含酒精漱口水以及用乙醇重构的无酒精漱口水时均表现出剂量依赖性的 DNA 损伤，如彗星试验所示。暴露于含酒精漱口水的转录组效应更为复杂，在发育不良（DOK）细胞中差异表达的基因数范围从>2000 个到正常（OKF6-TERT）细胞中<100 个。在 DOK 细胞中进行通路富集分析表明，两种使用的品牌的含酒精漱口水都具有共同的特征，包括 DNA 损伤反应和癌症相关通路。在 OKF6-TERT 细胞中，最显著富集的通路涉及炎症信号。
含酒精漱口水在体外对正常和发育不良的口腔角质细胞具有遗传毒性，并诱导广泛的基因表达变化。发育不良的细胞对漱口水的转录组效应更为敏感。
