Khan Nawal, Bavle Radhika M, Makarla Soumya, Konda Paremala, Amulya S R, Hosthor Sreenitha S
Department of Oral and Maxillofacial Pathology, Krishnadevaraya College of Dental Sciences, Bengaluru, Karnataka, India.
Department of Oral Pathology, Government Dental College and Hospital, Hyderabad, Telangana, India.
J Forensic Dent Sci. 2019 Sep-Dec;11(3):147-152. doi: 10.4103/jfo.jfds_29_19. Epub 2020 Jun 3.
DNA methylation (DNAm) age can be used to evaluate the chronological age of individuals often called "epigenetic age." In this study, buccal scrape samples were used for the determination of epigenetic age.
To examine if epigenetic age could be determined using neuronal pentraxin 2 (NPTX2) gene in buccal cells.
This cohort study was designed to validate the use of buccal cells for epigenetic age estimation. Sanger sequencing was used to determine the genetic sequence of the gene of interest postamplification. Nucleotide base sequence for NPTX2 gene was obtained for each case using this protocol.
The study was conducted on buccal scrapes obtained from 26 subjects of both genders, whose age varied from 1 to 65 years. The samples, collected by wooden spatulas, were placed in cell suspension buffer and stored at 4°C until transported to the laboratory.
Methylation levels of 5'-C-phosphate-G-3' located in the gene NPTX2 of 26 subjects were studied and analyzed by bisulfate sequencing. The percentage of methylation in this study falls in the range between 15% and 51%.
In this study, a sufficient amount of gDNA was retrieved from the buccal cells, thus confirming that buccal scrape was a feasible technique to obtain ample DNA. This study also showed that DNAm-polymerase chain reaction method was a feasible method for the evaluation of methylation pattern of NPTX2 gene.
DNA甲基化（DNAm）年龄可用于评估个体的实际年龄，通常称为“表观遗传年龄”。在本研究中，使用颊黏膜刮取样本测定表观遗传年龄。
研究是否可以使用颊细胞中的神经元五聚体蛋白2（NPTX2）基因来确定表观遗传年龄。
本队列研究旨在验证使用颊细胞进行表观遗传年龄估计的可行性。桑格测序用于在感兴趣基因扩增后确定其基因序列。使用该方案为每个病例获取NPTX2基因的核苷酸碱基序列。
本研究对26名年龄在1至65岁之间的男女颊黏膜刮取样本进行。样本由木质刮勺收集，置于细胞悬浮缓冲液中，在4°C下保存，直至运至实验室。
通过亚硫酸氢盐测序研究并分析了26名受试者NPTX2基因中位于5'-C-磷酸-G-3'处的甲基化水平。本研究中的甲基化百分比在15%至51%之间。
在本研究中，从颊细胞中获取了足够量的基因组DNA，从而证实颊黏膜刮取是获得充足DNA的可行技术。本研究还表明，DNAm聚合酶链反应方法是评估NPTX2基因甲基化模式的可行方法。
