利用细胞代谢组学探讨Nrf3在胃癌细胞增殖、迁移中的作用及机制  

To explore the role and mechanism of Nrf3 in the proliferation and migration of gastric cancer cells by cellular metabolomics

作　　者：陈梦未 陈雅慧 候维 胡清勇 周倩如 汤建才[1] CHEN Mengwei;CHEN Yahui;HOU Wei;HU Qingyong;ZHOU Qianru;TANG Jiancai(Institute of Basic Medicine and Forensics Medicine,North of Sichuan Medical College,Sichuan Nanchong 637000,China;Department of Pathology,North of Sichuan Medical College,Sichuan Nanchong 637000,China)

机构地区：[1]川北医学院基础医学与法医学院,四川南充637000 [2]川北医学院病理教研室,四川南充637000

出　　处：《现代肿瘤医学》2024年第7期1186-1193,共8页Journal of Modern Oncology

基　　金：四川省南充市科技局项目(编号:20SXCXTD004);川北医学院重点培育项目(编号:CBY22-ZDA-02)。

摘　　要：目的:探究核因子红系2-样3(NF-E2-related factor 3 or NFE2L3,Nrf3)对胃癌细胞代谢、增殖及迁移能力的影响及作用机制。方法:通过生物信息学分析Nrf3在胃癌组织中的表达;慢病毒构建基因沉默Nrf3的胃癌细胞株;基于人细胞UHPLC-OE-MS非靶标代谢组学技术,对胃癌细胞(AGS)及Nrf3敲降细胞进行代谢组学分析;利用RT-qPCR及Western blot实验检测敲降Nrf3对胆碱代谢关键酶的影响;采用CCK-8及划痕实验检测敲降Nrf3对胃癌细胞增殖、迁移的影响;通过外源性给予胆碱类似物氯化胆碱后观察胃癌细胞增殖、迁移能力的改变。结果:生物信息学分析发现胃癌组织中Nrf3表达高于癌旁组织;敲降Nrf3显著改变了胃癌细胞的代谢特征,鉴定到57个对分类有显著贡献的差异代谢物,其中32个代谢物在敲降Nrf3的胃癌细胞中减少,25个代谢物增加;Nrf3敲降导致胃癌细胞8条通路改变(P<0.05),其中胆碱、精氨酸和脯氨酸代谢改变最为明显(P<0.01);RT-qPCR及Western blot实验结果显示,敲降Nrf3后影响了胆碱代谢关键酶PLA2、LYPLA1、GPCPD1、CHK的表达,其中GPCPD1改变最为显著;CCK-8、划痕实验结果揭示了敲降Nrf3抑制胃癌细胞的增殖及迁移;外源性给予胆碱类似物氯化胆碱能部分逆转敲降Nrf3导致的胃癌细胞增殖、迁移能力的减弱。结论:敲降Nrf3抑制胃癌细胞增殖和迁移,其机制可能与胆碱代谢重编程有关。Objective:To investigate the effect of nuclear factor red lineage-like 3(NF-E2-related factor 3 or NFE2L3,Nrf3)on the metabolism,proliferation and migration of gastric cancer cells and its mechanism.Methods:The expression of Nrf3 in gastric cancer tissues was analyzed by bioinformatics.Lentivirus was constructed to silence Nrf3 gene in gastric cancer cell line.Based on human cell UHPLC-OE-MS non-target metabolomic technique,the metabolomic profiles of gastric cancer cells(AGS)and Nrf3 knock-down cells were analyzed.The effect of knockdown of Nrf3 on the key enzymes of choline metabolism was detected by RT-qPCR and Western blot.The effects of knockdown of Nrf3 on proliferation and migration of gastric cancer cells were examined by CCK-8 and scratch test.The changes of proliferation and migration of gastric cancer cells were observed after exogenous administration of choline analogue choline chloride.Results:The expression of Nrf3 in gastric carcinoma was higher than that in adjacent tissues by bioinformatics analysis.Knockdown of Nrf3 significantly altered the metabolic profile of gastric cancer cells,identifying 57 differential metabolites that contributed significantly to classification,of which 25 metabolites were decreased and 23 metabolites were increased in gastric cancer cells knockdown of Nrf3.Knockdown of Nrf3 resulted in the alteration of 8 pathways in gastric cancer cells(P<0.05),among which the changes of choline,arginine and proline metabolism were the most obvious(P<0.01).The results of RT-qPCR and Western blot showed that knockdown of Nrf3 affected the expression of PLA2,LYPLA1,GPCPD1 and CHK,especially GPCPD1.The results of CCK-8,and scratch experiments revealed that knockdown of Nrf3 inhibited the proliferation and migration of gastric cancer cells.Exogenous administration of cholinergic chloride partially reversed the inhibition of proliferation and migration induced by knockdown of Nrf3 in gastric cancer cells.Conclusion:Knockdown of Nrf3 inhibits the proliferation and migration of gastric cance

关 键 词：胃癌 Nrf3 胆碱代谢 增殖迁移 

分 类 号：R735.2[医药卫生—肿瘤]