摘要
目的:研究敲除芳香烃受体(aryl hydrocarbon receptor,AHR)基因对肝细胞癌(hepatocellular carcinoma,HCC)代谢途径和物质的影响。方法:利用成簇规律间隔短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)/Cas9技术构建AHR基因敲除的PLC/PRF/5肝癌细胞系,对野生型(AHR-WT)和AHR基因敲除(AHR-KO)肝癌细胞的代谢物质进行非靶向代谢组学检测,对其代谢物质差异进行分析,使用Metaboanalyst 4.0进行代谢通路富集分析,并寻找差异代谢物质。结果:将AHR-WT肝癌细胞和AHR-KO肝癌细胞分别进行正、负离子模式下的代谢物质检测,其前10位的功能富集结果包括氨基酸代谢(谷氨酸代谢、甘氨酸和丝氨酸代谢、精氨酸和脯氨酸代谢、蛋氨酸代谢、天冬氨酸代谢、尿素循环、氨再循环)、能量代谢[谷胱甘肽代谢、沃伯格(Warburg)效应]及核苷酸代谢(嘌呤代谢)。对差异代谢物质进行通路富集分析得到8条与肿瘤作用机制密切相关的代谢通路,包括精氨酸和脯氨酸代谢,精氨酸生物合成,嘧啶代谢,丙氨酸、天冬氨酸和谷氨酸代谢,苯丙氨酸、酪氨酸和色氨酸的生物合成,嘌呤代谢,赖氨酸退化,抗坏血酸和醛酸盐代谢。与AHR-WT肝癌细胞相比,AHR-KO肝癌细胞L-精氨酸、L-苯丙氨酸、L-酪氨酸、尿苷和黄嘌呤明显下调,而还原型烟酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide,NADH)、氧化型谷胱甘肽和还原型烟酰胺腺嘌呤二核苷酸磷酸(nicotinamide adenine dinucleotide phosphate,NADPH)明显上调。结论:AHR基因在HCC代谢过程中可能发挥重要作用,以AHR基因为靶点的HCC代谢组学变化可能是新的抗肿瘤治疗方案的研究方向。
Objective:To investigate the effect of aryl hydrocarbon receptor(AHR)gene knockout on metabolic pathways and metabolites in hepatocellular carcinoma(HCC).Method:PLC/PRF/5 hepatoma cell line with AHR knockout was constructed by clustered regularly interspaced short palindromic repeats(CRISPR)/Cas9 technology.Untargeted metabolomics was performed on the wild type(AHR-WT)and AHR knockout(AHR-KO)hepatoma cells to detect the metabolites,and the differential metabolites between the two groups were analyzed.Metaboanalyst 4.0 was used to perform metabolic pathway enrichment analysis and find differential metabolites.Result:The AHR-WT and AHR-KO hepatoma cells were examined for metabolites under positive and negative ionic model respectively.The results showed that the top 10 functional enrichments were mainly amino acid metabolism glutamate metabolism,glycine and serine metabolism,arginine and proline metabolism,methionine metabolism,aspartate metabolism,urea cycle,ammonia recycling,energy metabolism glutathione metabolism,Warburg effect and nucleotide metabolism purine metabolism.The pathway enrichment analysis showed that eight metabolic pathways were found including arginine and proline metabolism,arginine biosynthesis,pyrimidine metabolism,alanine,aspartic acid and glutamate metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,purine metabolism,lysine degradation,ascorbic acid and uronate metabolism.Compared with AHR-WT hepatoma cells,the levels of L-arginine,L-phenylalanine,L-tyrosine,uridine and xanthine were significantly down-regulated in AHR-KO hepatoma cells.Nicotinamide adenine dinucleotide(NADH),oxidized glutathione and nicotinamide adenine dinucleotide phosphate(NADPH)were significantly up-regulated in the AHR-KO cells.Conclusion:AHR may play an important role in the metabolism of HCC,and the research on the metabonomics of HCC by targeting AHR may be a new anti-tumor treatment strategy.
作者
魏哲文
朱青
蔡建强
Wei Zhewen;Zhu Qing;Cai Jianqiang(Department of Hepatobiliary Surgery,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital,Chinese Academy of Medical Science and Peking Union Medical College,Beijing 100021,China;Department of Clinical Laboratory Diagnostics,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)
出处
《肝癌电子杂志》
2023年第4期23-28,共6页
Electronic Journal of Liver Tumor
基金
国家自然科学基金青年科学基金(82302892)
中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-066)。
关键词
肝细胞癌
芳香烃受体
代谢组学
Hepatocellular carcinoma
Aryl hydrocarbon receptor
Metabolomics
