Zong Li, Pi Zifeng, Liu Shu, Xing Junpeng, Liu Zhiqiang, Song Fengrui
National Center of Mass Spectrometry in Changchun, Jilin Province Key Laboratory of Chinese Medicine Chemistry and Mass Spectrometry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China.
University of Chinese Academy of Sciences, Beijing, 100039, China.
Rapid Commun Mass Spectrom. 2018 Oct 15;32(19):1683-1692. doi: 10.1002/rcm.8229.
Multidrug resistance (MDR) occurs frequently and is a major challenge in tumor treatment. The lipid composition in the cell membrane and the redox balance are closely associated with the development of MDR. Liquid extraction surface analysis in combination with mass spectrometry (LESA-MS) has the characteristics of minimal sample preparation, rapid analysis, high sensitivity and high throughput, and has obtained wide applications.
LESA-MS was employed to in situ determine the lipids and other specific metabolites of intact MCF-7/ADR cells (adriamycin-resistant breast cancer cells) and its parental MCF-7/S cells grown on a glass slide. In situ atomic force microscopy was used to observe the morphology of tumor cells before and after extraction. Multivariate statistical analysis was used to investigate the potential lipid biomarkers correlated with the MDR. Moreover, the cell membrane fluidity and potential were determined.
The changes in the level of the lipids were closely correlated with the multidrug resistance of MCF-7/S cells. Moreover, lower cell membrane fluidity and higher cell membrane potential were observed and thus demonstrated the changes in the cell membrane induced by multidrug resistance. Also, the ratios of GSH/GSSG, ATP/ADP and ATP/AMP were significantly higher in MCF-7/ADR cells relative to MCF-7/S cells.
Lower cell membrane fluidity and higher cell membrane potential caused by the changes in lipid compositions, enhanced anti-oxidative ability and energy generation were involved in the development of the MDR. The specific alterations identified in this study may provide more information for overcoming MDR.
多药耐药（MDR）频繁发生，是肿瘤治疗中的一大挑战。细胞膜中的脂质组成与氧化还原平衡与MDR的发展密切相关。液相萃取表面分析结合质谱法（LESA-MS）具有样品制备最少、分析快速、灵敏度高和高通量的特点，并已得到广泛应用。
采用LESA-MS原位测定完整的MCF-7/ADR细胞（阿霉素耐药乳腺癌细胞）及其亲本MCF-7/S细胞在载玻片上生长时的脂质和其他特定代谢物。使用原位原子力显微镜观察提取前后肿瘤细胞的形态。采用多变量统计分析研究与MDR相关的潜在脂质生物标志物。此外，还测定了细胞膜流动性和电位。
脂质水平的变化与MCF-7/S细胞的多药耐药密切相关。此外，观察到较低的细胞膜流动性和较高的细胞膜电位，从而证明了多药耐药诱导的细胞膜变化。而且，相对于MCF-7/S细胞，MCF-7/ADR细胞中GSH/GSSG、ATP/ADP和ATP/AMP的比率显著更高。
脂质组成变化导致的较低细胞膜流动性和较高细胞膜电位、增强的抗氧化能力和能量生成参与了MDR的发展。本研究中确定的具体改变可能为克服MDR提供更多信息。
