胶质瘤条件培养基促进巨噬细胞糖酵解及表型转化  

Glioma conditioned medium promotes glycolysis and phenotypic transformation in macrophages

作　　者：张迪雅 李云帆 郭泓江 仇佳星 王钰铖 鞠瑞[1] 郭磊[1] ZHANG Diya;LI Yunfan;GUO Hongjiang;QIU Jiaxing;WANG Yucheng;JU Rui;GUO Lei(Department of Pharmacology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所、北京协和医学院基础学院药理系,北京100005

出　　处：《基础医学与临床》2024年第5期599-605,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(82002094);北京市自然科学基金(7222254);中国医学科学院医学与健康科技创新工程(2022-I2M-2-002)。

摘　　要：目的初步探索胶质瘤条件培养基诱导巨噬细胞糖酵解代谢与促癌介质表达的作用与机制。方法以经过缺氧与小鼠胶质瘤细胞系GL261条件培养基(GCM)诱导后的骨髓来源巨噬细胞(BMDM)(GCM-BMDM)为体外胶质瘤相关巨噬细胞(GAMs)模型,通过qPCR检测GCM-BMDM中M1与M2促癌介质、缺氧诱导因子-1α(HIF-1α)与糖酵解相关代谢酶基因mRNA水平;采用Seahorse细胞能量测定方法检测GCM-BMDM细胞外酸化速率(ECAR);通过Western blot检测信号传导与转录激活因子6(STAT6)、信号传导与转录激活因子3(STAT3)及其磷酸化形式p-STAT6、p-STAT3蛋白水平。结果GCM刺激BMDM Hif-1α与多种M1、M2促癌介质mRNA表达(P<0.01);GCM上调BMDM多种糖酵解基因mRNA表达(P<0.01),升高糖酵解水平(P<0.001),加入乳酸脱氢酶抑制剂stiripentol(STP)处理后糖酵解水平降低(P<0.05);STP下调GCM-BMDM Hif-1α与促癌介质Il-1β、Il-6、Ido与Cd163 mRNA水平(P<0.01);STP下调GCM-BMDM p-STAT3/STAT3与p-STAT6/STAT6比值(P<0.05)。结论GCM增加糖酵解代谢,促进HIF-1α、STAT3与STAT6介导的转录调控机制以诱导GAMs的促癌表型。STP能有效抑制GAMs糖酵解并降低GAMs促癌介质的转录表达水平。Objective To explore the effects and mechanisms involved in glioma conditioned medium for inducing glycolysis and oncogenic mediators in macrophages.Methods Bone marrow derived macrophage(BMDM)induced by hypoxia and mouse glioma cell line GL261 conditioned medium(GCM)(GCM-BMDM)were used as a model.The mRNA level of M1 and M2 oncogenic mediators,hypoxia-inducible factor-1α(HIF-1α)and glycolysis associated genes in GCM-BMDM were detected by qPCR.The extracellular acidification rate(ECAR)of GCM-BMDM was detected by Seahorse bioenergy assay method.The protein levels of signal transducer and activator of transcription 6(STAT6),signal transducer and activator of transcription 3(STAT3)and their phosphorylated forms p-STAT6 and p-STAT3 were examined by Western blot.Results GCM stimulated mRNA expression of Hif-1αand a variety of M1 and M2 oncogenic mediators in BMDM(P<0.01).GCM upregulated the mRNA expression of multiple glycolysis associated genes(P<0.01)and increased glycolysis level in BMDM(P<0.001).The glycolysis level was reduced by incubation with lactate dehydrogenase inhibitor stiripentol(STP)(P<0.05).STP down-regulated mRNA levels of Hif-1αand oncogenic mediators Il-1β,Il-6,Ido and Cd163 in GCM-BMDM(P<0.01).STP decreased the ratio of p-STAT3/STAT3 and p-STAT6/STAT6 in GCM-BMDM(P<0.05).Conclusions GCM increases glycolysis level and induces the pro-tumoral phenotypes in GAMs through promoting transcriptional regulation mediated by HIF-1α,STAT3 and STAT6.STP can effectively reduce glycolysis and transcriptional expression of oncogenic mediators in GAMs.

关 键 词：胶质瘤相关巨噬细胞 糖酵解 乳酸脱氢酶抑制剂 

分 类 号：R96[医药卫生—药理学]