四种肝（癌）细胞系糖代谢的流量解析

萨日娜; 武会娟

doi:10.3971/j.issn.1000-8578.2016.11.005

四种肝（癌）细胞系糖代谢的流量解析

Glucose Metabolic Flux of Four Liver (Hepatoma) Cell Lines

摘要

摘要:
目的分析人正常肝细胞（HL-7702）、不同转移潜能的肝细胞癌细胞系（SMMC-7721、HCCLM3）以及肝癌门静脉癌栓细胞系（CSQT2）的糖代谢流量及相关基因表达。
方法利用稳定同位素¹³C标记的葡萄糖与谷氨酰胺作为营养源，结合酶活实验，对四种细胞进行糖代谢流量解析。
结果 MTT与Transwell实验证实了HL-7702、SMMC-7721、HCCLM3、CSQT2细胞的增殖与迁移能力依次增加。酶活实验与标记实验结果均显示，糖酵解与三羧酸循环的代谢活性在四种细胞中依次增加，具体表现为葡萄糖消耗与乳酸产生的增加，代谢流量比率在多种糖代谢产物中的上升，尤其是在CSQT2细胞中达到最高。
结论随着肝癌细胞的恶性程度及转移能力的增加，糖酵解与三羧酸循环的代谢活性增加。代谢流量分析可能用以判断肝癌的发生发展与转移。

Abstract:
Objective To analyze glucose metabolic flux and related gene expression of human normal liver cells (HL-7702) ,hepatoma cell lines with different metastasis degree (SMMC-7721,HCCLM3) and hepatic portal vein tumor thrombus cell line CSQT2.
Methods Stable isotope ¹³C labeled glucose and glutamine were used as the nutrient source. Label assay and enzymes activity assay were employed to analyze glucose metabolic activity and flux of the four cell lines.
Results MTT and Transwell assay confirmed the proliferation and migration abilities were increased successively in HL-7702,SMMC-7721,HCCLM3 and CSQT2 cells. Label assay and enzyme activity assay showed that metabolic activity in glycolysis and tricarboxylic acid cycle(TCA cycle) was successively increased in the four cells,mainly reflected by the increase of glucose consumption and lactate production,as well as metabolic flux and ratio of glucose metabolism in hepatoma cell lines compared with normal liver cells,and reached the highest level in CSQT2 cell line.
Conclusion The metabolic pathway of glycolysis and TCA cycle is activated as malignant and metastasis degrees are increased in liver cancer cell lines. Metabolic flux analysis may help to diagnose the development and metastasis of liver cancer.

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