上海交通大学学报(医学版) ›› 2017, Vol. 37 ›› Issue (10): 1322-.doi: 10.3969/j.issn.1674-8115.2017.10.002?

• 论著(基础研究) • 上一篇    下一篇

乙型肝炎病毒影响肝癌HepG2细胞代谢的气相色谱-质谱研究

李静晨,万钱芬,干渺妍,唐惠儒   

  1. 复旦大学 生命科学学院,上海 200438
  • 出版日期:2017-10-28 发布日期:2017-11-01
  • 通讯作者: 唐惠儒,电子信箱:huiru_tang@fudan.edu.cn
  • 作者简介:李静晨(1991—),男,硕士生;电子信箱:14210700057@fudan.edu.cn
  • 基金资助:
     国家自然科学基金( 91439102,81590953)

HBV-induced metabolic changes in HepG2 human hepatoma cells revealed by gas chromatographymass spectrometry#br#

LI Jing-chen, WAN Qian-fen, GAN Miao-yan, TANG Hui-ru   

  1. School of Life Sciences, Fudan University, Shanghai 200438, China
  • Online:2017-10-28 Published:2017-11-01
  • Supported by:
    National Natural Science Foundation of China, 91439102, 81590953

摘要: 目的 · 研究乙型肝炎病毒(HBV) 感染对肝癌HepG2 细胞代谢的影响。方法 · 采用气相色谱– 质谱联用技术(GC-MS) 检测 肝癌HepG2 细胞和HepG2.2.15 细胞(HepG2 细胞的衍生细胞)的代谢物组成,分析其差异。结果 · GC-MS 分析发现HepG2 及 HepG2.2.15 细胞的亲水性提取物中主要含有 34 种代谢物,包括甘氨酸、丙氨酸、缬氨酸、亮氨酸、异亮氨酸、脯氨酸、丝氨酸、苏 氨酸、甲硫氨酸、半胱氨酸、胱氨酸、天冬氨酸、谷氨酸、焦谷氨酸、苯丙氨酸、酪氨酸、色氨酸、次黄嘌呤、尿嘧啶、肌醇、乳 酸、丁二酸、亚油酸、亚麻酸、软脂酸、硬脂酸、尿素和胆固醇等,覆盖糖酵解、脂肪酸代谢、氨基酸代谢、嘌呤与嘧啶代谢等多条 途径。HepG2.2.15 细胞中乳酸、亚麻酸、丙氨酸、半胱氨酸的含量显著低于HepG2 细胞,亮氨酸、异亮氨酸、缬氨酸、苯丙氨酸、 甲硫氨酸、色氨酸、脯氨酸、酪氨酸、肌醇、尿嘧啶等代谢物含量显著高于 HepG2 细胞。结论 · HBV 感染会导致 HepG2 细胞氨基酸 代谢和脂类代谢改变,基于 GC-MS 的代谢组学技术能为 HBV 感染的代谢应答机制提供补充信息。

关键词: 乙型肝炎病毒, HepG2 细胞, 代谢组学, 气相色谱 &ndash, 质谱联用技术

Abstract:

Objective · To investigate the impacts of hepatitis B virus (HBV) infection on the metabolomic phenotype of HepG2 human hepatoma cells.  Methods · With gas chromatography-mass spectrometry (GC-MS), metabolite composition of HepG2 and HepG2.2.15 cells (derived from HepG2 cells transfected with a plasmid containing HBV) were analysed.  Results · GC-MS analysis mainly found 34 metabolites in both HepG2 and HepG2.2.15 cells, including glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), serine (Ser), threonine (Thr), methionine (Met), cysteine (Cys), cystine, aspartic acid (Asp), glutamic acid (Glu), pyroglutamic acid, phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), hypoxanthine, uracil, myo-inositol, lactic acid, succinic acid, linoleic acid, linolenic acid, palmitic acid, stearic acid, urea, cholesterol, etc. These metabolites were involved in multiple metabolic pathways including glycolysis and metabolism of fatty acids, amino acids, purines and pyrimidines. Compared with HepG2 cells, HepG2.2.15 cells had significantly higher levels in lactic acid, linolenic acid, Ala and Cys, but lower levels in Leu, Ile, Val, Phe, Met, Trp, Pro, Tyr, myoinositol and uracil.  Conclusion · HBV infection dysregulates the metabolism of amino acids and fatty acids in hepatocytes. GC-MS analysis provides complimentary information about HBV-induced metabolic changes of host cells.

Key words:  hepatitis B virus, HepG2 cells, metabonomics, gas chromatography-mass spectrometry