您的位置:首页  > 论文页面

UPLC-MS/MS测定姜黄素在MCF-7/Adr细胞中的亚细胞分布研究

发表时间:2017-12-15  浏览量:1213  下载量:139
全部作者: 王金铃,李映,屠鹏飞
作者单位: 北京中医药大学中药学院
摘 要: 目的:研究姜黄素在阿霉素耐药乳腺癌(MCF-7/Adr)细胞中的亚细胞分布情况。方法:建立姜黄素超高效液相色谱串联质谱(ultra-high performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)测定方法,考察姜黄素溶液与MCF-7/Adr细胞共培养不同时间后,在MCF-7/Adr细胞不同亚细胞蛋白(细胞质、细胞膜/细胞器、细胞核和细胞骨架)中的分布情况。结果:姜黄素浓度在5~2 000 ng/mL浓度范围内线性良好,y=0.001 15x+0.005 17 (r=0.991 0);低、中、高3个浓度的平均回收率分别为99.21%、96.08%和100.45%,日间和日内精密度均低于15%. 姜黄素在0.5~2.0 h内,主要分布于细胞核、细胞膜/细胞器中,在细胞骨架和细胞质中少有分布。结论:定量评价了姜黄素在肿瘤细胞MCF-7/Adr细胞(细胞质、细胞膜/细胞器、细胞核和细胞骨架)中的定位及作用机制,为姜黄素抗肿瘤机制的研究提供了一定的实验依据。
关 键 词: 中药学;姜黄素;亚细胞分布;超高效液相色谱串联质谱(UPLC-MS/MS);阿霉素耐药乳腺癌细胞;蛋白沉淀
Title: Simultaneous determination of curcumin distribution in subcellular compartments of MCF-7/Adr cells by UPLC-MS/MS
Author: WANG Jinling, LI Ying, TU Pengfei
Organization: School of Chinese Materia Medica, Beijing University of Chinese Medicine
Abstract: Objective: To investigate distribution of curcumin in subcellular compartments of MCF-7/Adr cells. Methods: A specific and sensitive ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated for the determination of curcumin. After co-incubation with curcumin solution for different times, the distribution of curcumin was investigated in four MCF-7/Adr subcellular protein, including cytosolic, membrane/organelle, nucleus and cytoskeleton. Results: The calibration curves were linear with a concentration range of 5-2 000 ng/mL for curcumin (y=0.001 15x+0.005 17, r=0.991 0). The recoveries for the low, medium and high concentration samples were 99.21%, 96.08% and 100.45%, respectively. The inter-day and intra-day precisions were less than 15%. Curcumin were mainly distributed in nucleus and membrane/organelle during 0.5-2.0 h, seldom in cytoskeleton and cytosolic. Conclusion: We quantitatively determined the location and mechanism of curcumin in the MCF-7/Adr cells (such as cytosolic, membrane/organelle, nucleus and cytoskeleton). It will provide experimental data for the mechanism study of curcumin in antitumor.
Key words: science of Chinese pharmacology; curcumin; subcellular distribution; UPLC-MS/MS; MCF-7/Adr cells; protein precipitation
发表期数: 2017年12月第23期
引用格式: 王金铃,李映,屠鹏飞. UPLC-MS/MS测定姜黄素在MCF-7/Adr细胞中的亚细胞分布研究[J]. 中国科技论文在线精品论文,2017,10(23):2641-2648.
 
0 评论数 0
暂无评论
友情链接