Gaziantep University, Faculty of Dentistry, Department of Oral and Maxillofacial Surgery, Gaziantep, Turkey.
Bilkent University, Department of Molecular Biology and Genetics, Ankara, Turkey.
J Appl Oral Sci. 2020 Feb 7;28:e20190382. doi: 10.1590/1678-7757-2019-0382. eCollection 2020.
This study aimed to investigate the effects of Maras powder (a type of smokeless tobacco obtained from Nicotiana rustica Linn and mixed with the ashes of wood, especially from oak, walnut or grapevine) on the microRNA (miRNA) deregulation of oral mucosa, and it compares these effects with those of smoking.
Oral mucosal samples were collected from 74 patients, consisting of 16 nonusers, 26 smokers, and 32 Maras powder users. Genes associated with oral cancer were selected and 90 microRNAs targeting these genes were identified. MicroRNA were isolated and purified using the microRNA isolation kit. MicroRNA were expressed using Fluidigm RT-PCR.
A positive correlation between the duration of Maras powder use with miR-31 expression levels, and a negative correlation between the Maras powder chewing time and miR-372 expression levels was found. In addition, there is a negative correlation between the amount of Maras powder consumed and expression levels of miR-375, miR-378a, miR-145, and miR-10b; moreover, another negative correlation is observed between the number of cigarettes consumed and the expression levels of miR-23a, miR-23b, miR-203a, miR-200b, and miR-375. However, miR-200b and miR-92a levels were downregulated significantly more in Maras powder users when compared with smokers and nonusers (p<0.05).
The results show both chewing Maras powder and smoking have an effect on deregulation of miR-200b and miR-92a expressions. This leads to the belief that assessing the expression of these two miRNAs is a promising noninvasive method of analysis, especially in mutagen exposures. Finally, large-scale and high-throughput studies may help to identify an extensive miRNA expression profile associated with tobacco use and improve the understanding of oral malignancies.
本研究旨在探究玛拉斯粉（一种由菸草（Nicotiana rustica Linn）制成的无烟烟草，与木材灰分混合，特别是橡木、胡桃木或葡萄木灰分）对口腔黏膜中 microRNA（miRNA）失调的影响，并将其与吸烟的影响进行比较。
收集了 74 名患者的口腔黏膜样本，包括 16 名非使用者、26 名吸烟者和 32 名玛拉斯粉使用者。选择与口腔癌相关的基因，鉴定了 90 个靶向这些基因的 microRNA。使用 microRNA 分离试剂盒分离和纯化 microRNA。使用 Fluidigm RT-PCR 表达 microRNA。
发现玛拉斯粉使用时间与 miR-31 表达水平呈正相关，玛拉斯粉咀嚼时间与 miR-372 表达水平呈负相关。此外，玛拉斯粉的消耗量与 miR-375、miR-378a、miR-145 和 miR-10b 的表达水平呈负相关；此外，吸烟量与 miR-23a、miR-23b、miR-203a、miR-200b 和 miR-375 的表达水平也呈负相关。然而，与吸烟者和非使用者相比，玛拉斯粉使用者的 miR-200b 和 miR-92a 水平明显下调（p<0.05）。
结果表明，咀嚼玛拉斯粉和吸烟都会影响 miR-200b 和 miR-92a 的表达失调。这使得人们相信，评估这两个 miRNA 的表达是一种有前途的非侵入性分析方法，特别是在诱变剂暴露方面。最后，大规模和高通量的研究可能有助于确定与烟草使用相关的广泛 miRNA 表达谱，从而更好地了解口腔恶性肿瘤。
