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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：1876 下载量：236
| 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. |
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