鉴定与衰老相关的甲基化基因及筛选具有逆调控效应的物质

Identification of aging-related methylation genes and screening for substances with adverse regulatory effects

目的筛选和验证与人体衰老进程相关的甲基化基因,建立方便、灵敏、准确的检测方法,筛选能够逆向调控与衰老过程相关的甲基化基因的物质。方法通过GEO(Gene Expression Omnibus)公共数据库筛选并验证与人体衰老进程相关的甲基化基因,建立甲基化qPCR检测体系,验证目标基因与人体衰老的相关性。使用一组具有抗衰老功效的天然提取物/物质处理体外培养的细胞系,检测细胞系表型变化及目标基因甲基化水平变化。结果建立了促泌素(secretagogin,SCGN)和整合素亚基α2b(integrin subunit alpha 2b,ITGA2B)基因的甲基化检测体系,在53例独立样本中验证了SCGN和ITGA2B基因的甲基化调控与人体衰老进程相关。表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)和没食子酸(gallic acid,GA)抑制体外培养的细胞系的存活能力,且能逆转SCGN和ITGA2B基因在衰老进程中发生的甲基化调控。结论SCGN和ITGA2B基因甲基化水平的变化有可能用于评估人体衰老状态和监测人体衰老进程,同时也为开发基于DNA甲基化调控机制的抗衰老药物或产品提供了功效监测和评估工具。

Objective To screen and validate aging-associated DNA methylation dysregulation and to establish convenient,sensitive,accurate assays for detecting and screening substances capable of reversing dysregulated DNA methylation associated with the aging process.Methods The Gene Expression Omnibus(GEO)database was used to screen and validate DNA methylation alterations in the human aging process.A methylation qPCR detection system was developed to verify the correlation between DNA methylation levels of target genes and human age.Cancer cells were treated with a set of natural extracts/substances that had anti-aging properties to detect their effects on DNA methylation levels of target gene and phenotypic changes.Results A methylation detection system for the secretagogin(SCGN)and integrin subunit alpha 2b(ITGA2B)genes was established.Methylation levels of SCGN and ITGA2B genes were found to be correlated with human age,which was verified in 53 independent samples.Epigallocatechin gallate(EGCG)and gallic acid(GA)inhibited the tumor cell survival and reversed SCGN and ITGA2B methylation dysregulation associated with aging.Conclusions Measurements of SCGN a nd ITGA2B methylation level changes may be used to monitor the aging process in humans.They may also be used to provide experimental evidence for the development of anti-aging drugs or products based on DNA methylation regulation mechanisms.