基于代谢组学分析青蒿琥酯治疗肝癌的作用机制

王惠国; 王嗣瑶; 韩鑫龙; 李雨桐; 袁立霞; 谭晓梅; 汤庆发; 邢学锋; 陈飞龙; 尹雅蕾; 唐玲

doi:10.13422/j.cnki.syfjx.20191548

您当前的位置：

首页 >

文章列表页 >

基于代谢组学分析青蒿琥酯治疗肝癌的作用机制

药物代谢 | 更新时间：2021-02-09

- 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
- 中国实验方剂学杂志 2019年25卷第22期页码：78-85
- 作者机构：
  
  1.大连大学　生命科学与技术学院，辽宁　大连　 116622；
  2.中国科学院　大连化学物理研究所中国科学院分离分析化学重点实验室，转化医学科学研究中心，辽宁　大连　 116023；
  3.南方医科大学　中医药学院，广东省中药制剂重点实验室，广东省中药制剂技术工程实验室，广州　 510515
- 作者简介：
  
  王惠国，博士，副教授，从事天然产物活性成分分离及药理作用研究，Tel：0411-87402337，E-mail：903344836@qq.com
  尹雅蕾，博士，从事肿瘤耐药机制的研究，Tel：0411-84770960，E-mail：yinyalei1983@dicp.ac.cn
  唐玲，博士，教授，从事中药及复方药效物质基础及作用机制研究，Tel：020-62789112，E-mail：tangyuling6688@163.com；
- 基金信息：
  
  国家自然科学基金面上项目(81573661);广东省自然科学基金项目(2016A030313621)
- DOI：10.13422/j.cnki.syfjx.20191548
  中图分类号： R22; R945; R28; Q2; R969.1; O657
- 收稿日期：2018-12-25，
  
  网络出版日期：2019-04-09，
  
  纸质出版日期：2019-11-20
- 稿件说明：
移动端阅览
王惠国, 王嗣瑶, 韩鑫龙, 等. 基于代谢组学分析青蒿琥酯治疗肝癌的作用机制[J]. 中国实验方剂学杂志, 2019,25(22):78-85.

Hui-guo WANG, Si-yao WANG, Xin-long HAN, et al. Analysis of Mechanism of Artesunate in Treatment of Liver Cancer Based on Metabolomics[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(22): 78-85.
王惠国, 王嗣瑶, 韩鑫龙, 等. 基于代谢组学分析青蒿琥酯治疗肝癌的作用机制[J]. 中国实验方剂学杂志, 2019,25(22):78-85. DOI： 10.13422/j.cnki.syfjx.20191548.

Hui-guo WANG, Si-yao WANG, Xin-long HAN, et al. Analysis of Mechanism of Artesunate in Treatment of Liver Cancer Based on Metabolomics[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(22): 78-85. DOI： 10.13422/j.cnki.syfjx.20191548.

摘要

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(CellTiter-Glo® Luminescent Cell Viability Assay)检测不同浓度青蒿琥酯(0，12.5，25，50，100 μmol·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MetaboAnalyst 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(12.5～100 μmol·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。

Abstract

Objective:

To study on the antitumor mechanism of artesunate in the treatment of liver cancer based on gas chromatography-mass spectrometry (GC-MS).

Method:

CellTiter-Glo® Luminescent Cell Viability Assay was used to detect activity of artesunate with different concentrations (0

12.5

100 μmol·L

-1

) on human liver cancer Huh7

SMMC-7721 cells for 24

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 MetaboAnalyst 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 (12.5-100 μmol·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.

关键词

Keywords

references

Wisniewski J R , Vildhede A , Norén A , et al . In-depth quantitative analysis and comparison of the human hepatocyte and hepatoma cell line HepG2 proteomes [J]. J Proteomics , 2016 , 136 : 234 - 247 .

肖潺潺，陈茂剑，梅凡彪，等．辣椒碱对肝癌SMMC-7721细胞迁移和侵袭的影响及机制 [J]．中国实验方剂学杂志， 2018 ， 24 ( 18 )： 124 - 129 ．

Nzila A , Al-Zahrani I . Drugs for the treatment of malaria in the Kingdom of Saudi Arabia [J]. Saudi Med J , 2013 , 34 ( 6 ): 569 - 578 .

Karunajeewa H A , Kemiki A , Alpers M P , et al . Safety and therapeutic efficacy of artesunate suppositories for treatment of malaria in children in Papua New Guinea [J]. Pediatr Infect Dis J , 2003 , 22 ( 3 ): 251 - 256 .

Ilamathi M , Prabu P C , Ayyappa K A , et al . Artesunate obliterates experimental hepatocellular carcinoma in rats through suppression of IL-6-JAK-STAT signalling [J]. Biomed Pharmacother , 2016 , 82 : 72 - 79 .

QIAN P , ZHANG Y W , ZHOU Z H , et al . Artesunate enhances gammadelta T-cell-mediated antitumor activity through augmenting gammadelta T-cell function and reversing immune escape of HepG2 cells [J]. Immunopharmacol Immunotoxicol , 2018 , 40 ( 2 ): 107 - 116 .

JING W , DONG H , MIN M , et al . Dependence of artesunate on long noncoding RNA-RP11 to inhibit epithelial-mesenchymal transition of hepatocellular carcinoma [J]. J Cell Biochem , 2019 , 120 ( 4 ): 6026 - 6034 .

王高玉，刘红宁，戈淑超，等．铁皮石斛水提物对胃癌前病变作用的尿液代谢组学分析 [J]．中国实验方剂学杂志， 2018 ， 24 ( 21 )： 77 - 85 ．

Dunn W B , Broadhurst D I , Atherton H J , et al . Systems level studies of mammalian metabolomes: the roles of mass spectrometry and nuclear magnetic resonance spectroscopy [J]. Chem Soc Rev , 2011 , 40 ( 1 ): 387 - 426 .

刘欢，高洁，尹晶晶，等．不同防腐措施收集大鼠尿液对其内源性代谢物的影响 [J]．中国实验方剂学杂志， 2019 ， 25 ( 6 )： 120 - 124 ．

Pasikanti K K , Ho P C , CHAN E C . Gas chromatography/mass spectrometry in metabolic profiling of biological fluids [J]. J Chromatogr B Analyt Technol Biomed Life Sci , 2008 , 871 ( 2 ): 202 - 211 .

CHAN E C , Pasikanti K K , Nicholson J K . Global urinary metabolic profiling procedures using gas chromatography-mass spectrometry [J]. Nat Protoc , 2011 , 6 ( 10 ): 1483 - 1499 .

Lopes A S , Cruz E C , Sussulini A , et al . Metabolomic strategies involving mass spectrometry combined with liquid and gas chromatography [J]. Adv Exp Med Biol , 2017 , 965 : 77 - 98 .

Park H , Kim Y , Sul J W , et al . Synergistic anticancer efficacy of MEK inhibition and dual PI3K/mTOR inhibition in castration-resistant prostate cancer [J]. Prostate , 2015 , 75 ( 15 ): 1747 - 1759 .

王华，胡佳慧，刘翠钗，等．运用细胞代谢组学策略探究薯蓣皂苷元的抗肿瘤作用机制 [J]．中国实验方剂学杂志， 2018 ， 24 ( 20 )： 95 - 101 ．

ZHOU Y , SONG R , MA C , et al . Discovery and validation of potential urinary biomarkers for bladder cancer diagnosis using a pseudotargeted GC-MS metabolomics method [J]. Oncotarget , 2017 , 8 ( 13 ): 20719 - 20728 .

LI Z F , WANG J , HUANG C , et al . Gas chromatography/time-of-flight mass spectrometry-based metabonomics of hepatocarcinoma in rats with lung metastasis: elucidation of the metabolic characteristics of hepatocarcinoma at formation and metastasis [J]. Rapid Commun Mass Spectrom , 2010 , 24 ( 18 ): 2765 - 2775 .

WANG Y , YAN L , LIU J , et al . The HNF1α-regulated lncRNA HNF1α-AS1 is involved in the regulation of cytochrome P450 expression in human liver tissues and Huh7 cells [J]. J Pharmacol Exp Ther , 2019 , 368 ( 3 ): 353 - 362 .

CHEN W , WU J , SHI H , et al . Hepatic stellate cell coculture enables sorafenib resistance in Huh7 cells through HGF/c-Met/Akt and Jak2/Stat3 pathways [J]. Biomed Res Int , 2014 , doi: 10.115/2014/764981 http://doi.org/10.115/2014/764981 .

Kwon S , Jeon J S , Ahn C , et al . Rapamycin regulates the proliferation of Huh7, a hepatocellular carcinoma cell line, by up-regulating p53 expression [J]. Biochem Biophys Res Commun , 2016 , 479 ( 1 ): 74 - 79 .

Halama A , Möller G , Adamski J . Metabolic signatures in apoptotic human cancer cell lines [J]. OMICS , 2011 , 15 ( 5 ): 325 - 335 .

Nevedomskaya E , Ramautar R , Derks R , et al . CE-MS for metabolic profiling of volume-limited urine samples: application to accelerated aging TTD mice [J]. J Proteome Res , 2010 , 9 ( 9 ): 4869 - 4874 .

杨婧，贾彦，王蔚，等．膈下逐瘀汤对大鼠纤维化肝脏组织谷胱甘肽抗氧化系统的影响 [J]．中国实验方剂学杂志， 2017 ， 23 ( 17 )： 121 - 126 ．

Spitzer S , Volbracht K , Lundgaard I , et al . Glutamate signalling: a multifaceted modulator of oligodendrocyte lineage cells in health and disease [J]. Neuropharmacology , 2016 , 110 ( Pt B ): 574 - 585 .

Jain M , Nilsson R , Sharma S , et al . Metabolite profiling identifies a key role for glycine in rapid cancer cell proliferation [J]. Science , 2012 , 336 ( 6084 ): 1040 - 1044 .

Sreekumar A , Poisson L M , Rajendiran T M , et al . Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression [J]. Nature , 2009 , 457 ( 7231 ): 910 - 914 .

ZHANG W C , Shyh-Chang N , YANG H , et al . Glycine decarboxylase activity drives non-small cell lung cancer tumor-initiating cells and tumorigenesis [J]. Cell , 2012 , 148 ( 1/2 ): 259 - 272 .

Possemato R , Marks K M , Shaul Y D , et al . Functional genomics reveal that the serine synthesis pathway is essential in breast cancer [J]. Nature , 2011 , 476 ( 7360 ): 346 - 350 .

WANG J , Alexander P , WU L , et al . Dependence of mouse embryonic stem cells on threonine catabolism [J]. Science , 2009 , 325 ( 5939 ): 435 - 439 .

CHEN C Y , Hsu Y L , CHEN Y Y , et al . Isokotomolide A, a new butanolide extracted from the leaves of Cinnamomum kotoense, arrests cell cycle progression and induces apoptosis through the induction of p53/p21 and the initiation of mitochondrial system in human non-small cell lung cancer A549 cells [J]. Eur J Pharmacol , 2007 , 574 ( 2/3 ): 94 - 102 .

Feun L , YOU M , WU C J , et al . Arginine deprivation as a targeted therapy for cancer [J]. Curr Pharm Des , 2008 , 14 ( 11 ): 1049 - 1057 .

Phang J M , Pandhare J , LIU Y . The metabolism of proline as microenvironmental stress substrate [J]. J Nutr , 2008 , 138 ( 10 ): 2008S - 2015S .

Watford M . Glutamine metabolism and function in relation to proline synthesis and the safety of glutamine and proline supplementation [J]. J Nutr , 2008 , 138 ( 10 ): 2003S - 2007S .

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

基于UPLC-Q-TOF-MS/MS分析粉葛芦头的化学成分及其对小鼠肝脏代谢物的影响

基于血清代谢组学探讨青翘、老翘对急性肺损伤大鼠的作用机制

基于代谢组学探讨柴芍六君汤对肝郁脾虚型慢性萎缩性胃炎大鼠的中枢调节作用

基于¹H-NMR代谢组学的豨莶草水洗脱部位致大鼠肺损伤作用机制分析

独活续断汤对肝肾阴虚型腰间盘突出症患者临床疗效及血清代谢组学的影响