基于代谢组学分析青蒿琥酯治疗肝癌的作用机制

Analysis of Mechanism of Artesunate in Treatment of Liver Cancer Based on Metabolomics

目的:采用GC-MS联用技术探索青蒿琥酯治疗肝癌的作用机制,为该药物在肝癌治疗中的临床应用提供实验依据。方法:应用发光法细胞活力检测试剂盒(Cell Titer-Glo? Luminescent Cell Viability Assay)检测不同浓度青蒿琥酯(0,12. 5,25,50,100μmol·L-1)作用于人肝癌Huh7,SMMC-7721细胞24,48,72 h的活性。采用GC-MS联用技术分析青蒿琥酯作用于2种肝癌细胞(Huh7,SMMC-7721) 24 h后代谢物的变化,利用Postrun Analysis 4. 41工作站对数据进行预处理,采用偏最小二乘法-判别分析(PLS-DA)对两组差异代谢物进行分析,通过Metabo Analyst 3. 0软件对差异代谢物进行代谢通路分析。结果:与正常组相比,青蒿琥酯作用于2种肝癌细胞后,细胞内相同代谢物共有39种发生了显著变化,主要涉及到氨基酰-转运RNA(tRNA)生物合成,丙氨酸、天冬氨酸和谷氨酸代谢,甘氨酸、丝氨酸和苏氨酸代谢,精氨酸和脯氨酸代谢,谷胱甘肽代谢5条代谢通路。结论:青蒿琥酯(12. 5~100μmol·L-1)均能够抑制肝癌细胞(Huh7,SMMC-7721)的生长,主要涉及到5条代谢通路,这可能是青蒿琥酯抗肝癌的作用途径。

Objective:To study on the antitumor mechanism of artesunate in the treatment of liver cancer based on gas chromatography-mass spectrometry(GC-MS).Method:Cell Titer-Glo?Luminescent Cell Viability Assay was used to detect activity of artesunate with different concentrations(0,12.5,25,50,100μmol·L-1)on human liver cancer Huh7,SMMC-7721 cells for 24,48,72 h.GC-MS was employed to analyze the changes of metabolites of artesunate in two kinds of hepatoma cells(Huh7,SMMC-7721)for 24 h.The data was preprocessed by Postrun Analysis 4.41 workstation.Partial least squares-discriminant analysis(PLS-DA)was used to analyze two sets of differential metabolites and to analyze metabolic pathways of differential metabolites based on Metabo Analyst 3.0 software.Result:Compared with the normal group,after two kinds of liver cancer cells was treated by artesunate,a total of 39 identical metabolites in the cells have undergone significant changes,which were mainly related to five metabolic pathways,including biosynthesis of aminoacyl-transfer RNA(tRNA),metabolism of alanine,aspartic acid and glutamic acid,metabolism of glycine,serine and threonine,metabolism of arginine and proline,metabolism of glutathione.Conclusion:Artesunate(12.5-100μmol·L-1)can inhibit the growth of liver cancer cells(Huh7,SMMC-7721),it mainly involves five metabolic pathways,which may be the pathway of artesunate against liver cancer.