摘要
建立了一种基于超高效液相色谱-离子阱-飞行时间质谱(UPLC-IT-TOF MS)联用的非靶标代谢组学方法,研究了人慢性髓系白血病细胞及其阿霉素耐药细胞(K562和K562-ADM)之间的代谢物差异。考察了细胞破碎方法、提取溶剂体系和溶剂体积对细胞代谢物提取效果的影响,实验发现采用超声破碎法,以800μL的80%甲醇提取时可得到较多的代谢特征峰,方法重复性和稳定性较高。将建立的方法应用于K562及K562-ADM细胞的代谢组差异研究,对所得数据进行主成分分析(PCA)和正交偏最小二乘法判别分析(OPLS-DA)。结果显示2种细胞的代谢特征存在显著差异,并发现40个对分类有显著贡献的代谢物,差异代谢物主要参与氨基酸代谢、鞘脂代谢、嘌呤代谢和嘧啶代谢等通路。该方法可用于K562和K562-ADM细胞的代谢组差异研究,可望为白血病细胞耐药性的代谢组学研究提供帮助。
Based on ultra performance liquid chromatography coupled with ion trap-time-of-flight mass spectrometry(UPLC-IT-TOF MS),a non-targeted metabolomics method was developed for the investigation on metabolite difference between human chronic myeloid leukemia cells and their adriamycin-resistant cells,i.e.K562 and K562-ADM.Specifically,impacts of disruption methods,extraction solvent systems and volumes on the extraction effects for metabolites in QC samples were examined.Results showed that the ultrasonic disruption method in an 800μL of 80%methanol solvent system could obtain more feature peaks from leukemia cells with good repeatability and stability.Then,the established method was applied to investigate the metabolite difference between K562 and K562-ADM.The data was processed by principal component analysis(PCA)and orthogonal partial least squares discrimination analysis(OPLS-DA),and the significant differences between the metabolome of K562 and that of K562-ADM cells were obtained,in which forty metabolites significantly contributed to the differences,and they were mainly involved in amino acid metabolism,sphingolipid metabolism,purine metabolism and pyrimidine metabolism.The established method could be used to investigate the metabolite differences between K562 and K562-ADM,and further provide some help for future metabolomics researches on drug-resistance of leukemia cells.
作者
周玲婳
卢红梅
ZHOU Ling-hua;LU Hong-mei(College of Chemistry and Chemical Engineering,Central South University,Changsha 410083,China)
出处
《分析测试学报》
CAS
CSCD
北大核心
2020年第7期821-828,共8页
Journal of Instrumental Analysis
基金
国家自然科学基金(21873116,21675174)。
关键词
白血病细胞
非靶标代谢组学
超高效液相色谱-质谱联用
阿霉素耐药性
leukemia cells
non-targeted metabolomics
ultra performance liquid chromatography-tandem mass spectrometry
adriamycin resistance
