乙型肝炎病毒影响肝癌HepG2细胞代谢的气相色谱-质谱研究

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

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。

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.