细胞代谢组学用于羽扇豆醇干预人乳腺癌细胞MCF-7的机理探究

Mechanism research on the lupeol treatment on MCF-7 breast cancer cells based on cell metabonomics

应用基于气相色谱-质谱联用(GC-MS)的代谢组学方法结合细胞周期实验,研究羽扇豆醇体外抑制人乳腺癌细胞MCF-7增殖的作用机理。代谢组学的研究结果表明:通过正交偏最小方差判别分析(OPLS-DA)可以很好地区分羽扇豆醇作用的MCF-7细胞代谢谱与对照组细胞代谢谱,模型参数为:R2 Ycum=0.988,Q2 Ycum=0.964。VIP(variable importance in the projection)值大于1的差异代谢物进一步用t检验进行单位分析,选择t<0.05(VIP>1)的代谢物作为羽扇豆醇作用组的生物标志物,得到琥珀酸、磷酸、亮氨酸、异亮氨酸等11种代谢差异物。结合羽扇豆醇将细胞周期抑制在G1期这一现象,推测羽扇豆醇可能是主要抑制了三羧酸循环中的琥珀酰辅酶A的生成和底物磷酸化生成ATP的反应来抑制MCF-7细胞的增殖。本实验从代谢组学角度为乳腺癌抗肿瘤机制提供新的线索。

The objective of this research is to investigate the suppressive effects of lupeol on MCF-7 breast cancer cells,and explore its mechanism on inhibiting the proliferation of MCF-7 cells based on cell metabonomics and cell cycle. Gas chromatography-mass spectrometry（GC-MS）was used in the cell metabonomics assay to identify metabolites of MCF-7 cells and MCF-7 cells treated with lupeol. Then,orthogonal partial least squares discriminant analysis（OPLS-DA）was used to process the metabolic data and model parameters of OPLS-DA were as fol-lows：R2 Ycum = 0. 988,Q 2 Ycum = 0. 964,which indicated that these two groups could be distin-guished clearly. The metabolites（ VIP（ variable importance in the projection）﹥ 1）were ana-lyzed by t-test,and finally,metabolites（ t﹤0. 05）were identified to be biomarkers. Eleven metabolites such as butanedioic acid,phosphoric acid,L-leucine and isoleucine which had a significant contribution to classification were selected and preliminarily identified due to the accurate mass. Cell cycle assay was analyzed by FACSCalibur. Since the cells in the phase of G1 were increased significantly after the treatment of lupeol,we speculated that lupeol has a bloc-king effect on the generation of succinyl-CoA and the reaction of substrate phosphorylation of tricarboxylic acid cycle of MCF-7 cells. This study provided a novel approach to the mechanism research on the lupeol treatment on MCF-7 breast cancer cells based on cell metabonomics.