南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (12): 1765-1773.doi: 10.12122/j.issn.1673-4254.2022.12.03

• • 上一篇    下一篇

NDRG2通过调控肝癌细胞磷脂和甘油三酯代谢抑制肝细胞癌的生长：基于代谢组学分析

王佳媛，袁依依，张 坤，孙 翔，卜 歆，董 健，吴有盛，田红英，沈 岚   

  1. 延安大学医学院病原生物学教研室，陕西 延安 716000；空军军医大学基础医学院生物化学与分子生物学教研室，口腔医学院，陕西 西安 710032
  • 出版日期:2022-12-20 发布日期:2023-01-12

NDRG2 inhibits tumorigenesis of hepatocellular carcinoma by regulating metabolism of phospholipids and triglyceride: a metabonomic analysis

WANG Jiayuan, YUAN Yiyi, ZHANG Kun, SUN Xiang, BU Xin, DONG Jian, WU Yousheng, TIAN Hongying, SHEN Lan   

  1. Department of Pathogenic Biology, Medical College, Yan'an University, Yan'an 716000, China; State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, School of Stomatology, Air Force Military Medical University, Xi'an 710032, China
  • Online:2022-12-20 Published:2023-01-12

RichHTML

34

PDF

344

摘要：

目的 观察抑癌基因N-Myc下游调控基因2（NDRG2）对人肝癌细胞脂类物质代谢的调控方式及调控特征，阐述NDRG2在肝癌脂质代谢中的作用。方法 基于TNMplot数据库分析抑癌基因NDRG2在809例肝细胞癌组织和379例正常肝组织中的表达差异，应用人类蛋白质图谱（THPA）分析NDRG2在肝细胞癌的表达水平与患者生存期的关系，以及NDRG2在肿瘤细胞株的表达丰度并筛选肝癌细胞系。利用携带NDRG2 cDNA慢病毒及其对照组慢病毒分别感染人肝细胞癌细胞系HepG2，采用实时荧光定量PCR法检测慢病毒感染后两组细胞中NDRG2基因的mRNA含量；Western blot检测慢病毒感染后两组细胞中NDRG2的蛋白含量，建立NDRG2过表达及其对照组细胞系。利用脂质代谢组学分析NDRG2表达增强对肝癌细胞脂质代谢的调控效应，利用酶联免疫分析试剂盒验证NDRG2表达增强对肝癌细胞磷脂代谢物的影响，利用油红染色验证NDRG2表达增强对肝癌细胞甘油三酯的影响。结果 TNMplot数据库结果统计分析发现NDRG2在肝癌组织中表达量对比普通肝组织明显下降（P<0.001）；THPA数据库生存分析发现NDRG2 mRNA水平高的患者比NDRG2 mRNA水平低的患者存活时间更长；THPA数据库细胞系RNA表达分析显示在多种肿瘤细胞中，肝细胞癌HepG2中NDRG2表达丰度较高。代谢组学研究发现NDRG2表达增强能够显著调控肝癌细胞HepG2磷脂的含量。其中甘油三酯TG、卵磷脂PC、磷脂酰甘油PG、磷脂酰乙醇胺PE、鞘磷脂酰丝氨酸SM、神经酰胺Cer等变化明显。酶联免疫分析发现NDRG2表达增强能够显著抑制肝癌细胞HepG2内多种磷脂代谢物的含量。油红染色分析发现NDRG2表达增强能够显著抑制肝癌细胞HepG2内甘油三酯的含量。结论 抑癌基因NDRG2可以通过调控肝癌细胞磷脂和甘油三酯含量，抑制肝细胞癌的发生与发展。

关键词: 抑癌基因；NDRG2；磷脂；代谢组学

Abstract: Objective To explore the role of the tumor suppressor gene NDRG2 in regulating lipid metabolism in hepatoma cells. Methods We analyzed the differential expression of NDRG2 gene between hepatocellular carcinoma tissues (n=809) and normal liver tissues (n=379) based on data from TNMplot database, and investigated the correlation between NDRG2 mRNA expression and the overall survival of the patients with hepatocellular carcinoma using THPA database, which was also used for analysis of NDRG2 expression levels in tumor cell lines for screening hepatoma cell lines. Human hepatoma cell line HepG2 was infected with a lentivirus containing NDRG2 cDNA, and the expression level of NDRG2 in the infected cells was detected using qPCR and Western blotting. Lipid metabolomics analysis was performed to analyze the regulatory effect of NDRG2 overexpression on lipid metabolism in HepG2 cells, and ELISA and Oil Red O staining were used to examine the changes in contents of phospholipids and triglyceride in NDRG2-overexpressing HepG2 cells. Results Analysis of the TNMplot database showed that NDRG2 expression level was significantly lower in hepatocellular carcinoma tissues than in normal liver tissues (P<0.001). Analysis of THPA database showed that the patients with high NDRG2 mRNA levels had a longer survival time than those with low NDRG2 mRNA levels, and NDRG2 expression level was the highest in HepG2 cell line among the tumor cell lines. Metabolomics analysis showed that in HepG2 cells, NDRG2 overexpression led to changes in the contents of phospholipids, and among them lecithin PC, phosphatidyl glycerol PG, phosphatidyl ethanolamine PE, sphinophosphatidyl serine SM, and ceramide Cer exhibited significant changes. The results of ELISA and Oil Red O staining demonstrated that NDRG2 overexpression obviously reduced the contents of multiple phospholipids and significantly lowered the contents of triglyceride in HepG2 cells. Conclusion NDRG2 regulates tumorigenesis of hepatocellular carcinoma by modulating the metabolism of phospholipids and triglyceride.

Key words: tumor suppressor gene; NDRG2; phospholipids; metabolomics