干扰素-γ刺激人巨噬细胞后的转录组变化与细胞内信号调控机制研究  

Transcriptional Modification and Potential Intracellular Signaling Mechanisms in Human Macrophages Primed by Interferon-γ

作　　者：刘蓓 高泓浩 程莉 张家乐 董焱鑫 谢顺 黄文荣 袁顺宗 LIU Bei;GAO Hong-Hao;CHENG Li;ZHANG Jia-Le;DONG Yan-Xin;XIE Shun;HUANG Wen-Rong;YUAN Shun-Zong(The PLA 307 Clinical College of Anhui Medical University,Beijing 100071,China;Section 4,Department of Hematology,The Fifh Medical Center of PLA General Hospital,Beijing 100071,China;Department of Thoracic Surgery,The Sixth Medical Center of PLA General Hospital,Beijing100048,China)

机构地区：[1]安徽医科大学解放军307临床学院,北京100071 [2]解放军总医院第五医学中心血液病医学部血液科四病区,北京100071 [3]解放军总医院第六医学中心胸外科,北京100048

出　　处：《中国实验血液学杂志》2022年第5期1590-1595,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金(81770207,82170230);科技部国家重点研发计划(2016YFC1101102)。

摘　　要：目的:探讨干扰素-γ(IFN-γ)刺激后人巨噬细胞内上调的转录组基因表达谱及其相应的细胞内信号调控机制。方法:运用RNA-seq对IFN-γ刺激前后的人巨噬细胞系U937差异基因表达谱进行测序和比较,从中筛选出显著上调的基因,利用实时荧光定量PCR技术(qPCR)分别在U937和THP1两个巨噬细胞株内进行验证;在IFN-γ刺激U937细胞的同时,分别加入JAK/STAT3、MAPK/ERK和PI3K/AKT通路抑制剂,检测其对上调基因的影响,从中找出关键的调控机制。结果:巨噬细胞经IFN-γ刺激后,RNA-seq筛选结合qPCR验证结果显示:趋化因子CXCL9、CXCL10、CXCL11,APOL家族包括APOL1、APOL2、APOL3、APOL4、APOL6以及GBP家族GBP1、GBP2、GBP3、GBP4、GBP5均显著上调。JAK/STAT3通路抑制剂抑制了IFN-γ诱导的APOL1、APOL4、GBP1、GBP4和GBP5基因上调表达,MAPK/ERK通路抑制剂抑制了IFN-γ诱导的CXCL10基因上调表达,PI3K/AKT通路抑制剂抑制了IFN-γ诱导的APOL1、APOL4、APOL6、GBP1和GBP5基因上调表达,三种信号通路抑制剂均能抑制IFN-γ诱导的CXCL9基因上调表达,三种通路抑制剂对IFN-γ诱导的APOL3基因上调均无抑制作用。结论:IFN-γ刺激后巨噬细胞内APOL和GBP部分家族分子表现为显著上调,PI3K/AKT、JAK/STAT3和MAPK/ERK通路对其表达分别具有一定的正向调控作用。Objective:To explore the transcriptional gene expression profile up-regulated in human macrophages stimulated by interferon-γ(IFN-γ)and the underlying intracellular signaling mechanisms. Methods:RNA-seq was used to sequence and compare the differential gene expression profiles of human macrophage cell line U937before and after IFN-γstimulation,and the significantly up-regulated genes were screened out,which were verified by fluorescence-based real-time quantitative polymerase chain reaction(qPCR)in U937and THP1cell lines,respectively. JAK/STAT,MAPK/ERK and PI3K/AKT pathway inhibitors were added to simultaneously to the cultured U937cells upon IFN-γpriming to detect their effects on the expressions of the up-regulated genes to explore the key regulatory mechanisms.Results:RNA-seq and qPCR results showed that,the well-recognized chemokines CXCL9,CXCL10 and CXCL11,the APOL family including APOL1,APOL2,APOL3,APOL4,APOL6 and GBP family GBP1,GBP2,GBP3,GBP4 and GBP5 as well were significantly up-regulated in IFN-γ-stimulated U937cells. JAK/STAT3pathway inhibitor inhibited the upregulation of APOL1,APOL4,GBP1,GBP4 and GBP5 genes induced by IFN-γ,while MAPK/ERK pathway inhibitor inhibited the upregulation of CXCL10 gene. PI3K/AKT pathway inhibitor inhibited the upregulation of APOL1,APOL4,APOL6,GBP1 and GBP5 genes induced by IFN-γ,all three signal pathway inhibitors could inhibit the upregulation of CXCL9 gene,and none of them could inhibit the upregulation of APOL3 gene. Conclusion:Upon IFN-γstimulation,some family molecules of APOL and GBP in macrophages are significantly up-regulated,and PI3K/AKT,JAK/STAT3and MAPK/ERK pathways have positive regulation on their expressions,respectively.

关 键 词：干扰素-Γ 人巨噬细胞 转录组 细胞内信号 

分 类 号：R392[医药卫生—免疫学] R551[医药卫生—基础医学]