常氧和低氧条件下二甲双胍对脑胶质瘤细胞代谢的影响  

The effect of metformin on glioma cell metabolism under normoxic and hypoxic conditions

作　　者：梅蕾 汤旗 厉柳岑 段荣清 沈耀[1] MEI Lei;TANG Qi;LI Liucen;DUAN Rongqing;SHEN Yao(College of Life Sciences&Laboratory Medicine,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学检验医学院(生命科学学院),浙江温州325035

出　　处：《温州医科大学学报》2023年第6期440-449,共10页Journal of Wenzhou Medical University

基　　金：浙江省自然科学基金项目(LY19H090010);温州医科大学基本科研业务项目(KYYW201903)。

摘　　要：目的:探索在常氧和低氧条件下二甲双胍对脑胶质瘤U251细胞代谢的影响。方法:在常氧(21%O2)和低氧(1%O2)条件下,使用25 mmol/L的二甲双胍在处理脑胶质瘤U251细胞48 h后,采用MTT法检测U251细胞的细胞活力,平板克隆法检测细胞形成克隆的能力,EdU法检测细胞的DNA增殖变化,细胞周期试剂盒检测细胞的周期变化,超高效液相色谱-高分辨质谱非靶向代谢检测法检测细胞的代谢变化,ATP和乳酸试剂盒分别检测细胞内的ATP含量和胞外乳酸的含量,Western blot检测葡萄糖转运蛋白(GLUT1)和原癌基因(c-Myc)蛋白的相对表达量。结果:在常氧和低氧条件下,25 mmol/L二甲双胍处理脑胶质瘤U251细胞48 h后,与对照组相比,细胞的活力、细胞形成克隆率、细胞增殖率以及细胞代谢物含量均出现降低(P<0.05)。常氧条件下,二甲双胍增加了脑胶质瘤U251细胞的ATP含量和胞外乳酸含量(P<0.05);而低氧条件下,经二甲双胍处理后脑胶质瘤U251细胞内的GLUT1和c-Myc蛋白的相对表达量显著下降(P<0.05)。结论:二甲双胍通过下调脑胶质瘤U251细胞的代谢来抑制脑胶质瘤U251细胞的增殖和形成克隆的能力。Objective:To explore the effect of metformin on the metabolism of glioma U251 cells under normoxic and hypoxic conditions.Methods:Under the conditions of normox(21%O2)and hypoxia(1%O2),metformin at 25 mmol/L was used to detect the cell viability of U251 cells by MTT method;the ability of cells to form clones by plate cloning method;the DNA proliferation changes of cells were detected by EdU method;the cycle changes of cells were detected by cell cycle kits;the metabolic changes of cells were detected by ultra-high performance liquid chroma-high-resolution mass spectrometry untargeted metabolism detection.The content of ATP in cells and the content of extracellular lactic acid were detected by ATP and lactate kits,and the relative expression of glucose transporter(GLUT1)and proto-oncogene(c-Myc)proteins were detected by Western blot method,respectively.Results:Compared with the control group,the viability,cell formation cloning rate,cell proliferation rate and cell metabolite content were reduced(P<0.05)after 25 mmol/L metformin treated with glioma U251 cells for 48 h under normal oxygen and hypoxic conditions.Under normal oxygen conditions,metformin increased the ATP content and extracellular lactate content of brain glioma U251 cells(P<0.05).The relative expression of GLUT1 and c-Myc proteins in U251 cells after metformin treatment under hypoxic conditions decreased significantly(P<0.05).Conclusion:Metformin inhibits the proliferation of glioma U251 cells and the ability to form clones by downregulating the metabolism of glioma U251 cells.

关 键 词：低氧 二甲双胍 U251细胞 细胞代谢 细胞增殖 

分 类 号：R730.5[医药卫生—肿瘤]