慢性粒细胞白血病T细胞TCRζ链表达上调后全基因表达谱改变的分析  

The Genome-Wide Changes in Expression Profile of CML T Cells After Up-regulation of TCRζ Chain Expression

作　　者：黎维丹 练佳仪 陈少华 李扬秋 查显丰[1] LI Wei-Dan;LIAN Jia-Yi;CHEN Shao-Hua;LI Yang-Qiu;ZHA Xian-Feng(Department of Clinical Laboratory,School of Medicine,First Affiluited Hospital,Jinan University;Institute of Hematology,Medical College,Jinan University,Guanfizhou 510632,Guangdong Province,China)

机构地区：[1]暨南大学附属第一医院临床医学检验中心 [2]暨南大学基础医学院血液病研究所,广东广州510632

出　　处：《中国实验血液学杂志》2021年第3期669-676,共8页Journal of Experimental Hematology

基　　金：国家自然科学基金青年基金项目(81400109,81770152);广东省自然科学基金博士启动项目(S2013040016151)。

摘　　要：目的:分析表达上调TCRζ后慢性粒细胞白血病(CML)患者T细胞基因表达谱的变化特点,探讨转基因上调TCRζ后T细胞活性恢复的分子机制。方法:收集3例初发未治疗慢性期CML患者的外周血单个核细胞(PBMCs),利用MACS法分选获得CD3+T细胞;利用核转染技术转染TCRζ-IRES2-EGFP(实验组)和pIRES2-EGFP(对照组)质粒至T细胞中;应用AffymetrixGeneChip Human Gene 2.0 ST Array芯片对上调TCRζ链表达的CML T细胞和对照细胞基因表达谱进行检测,并对差异表达基因进行GO功能注释分析和KEGG通路的富集分析。结果:共筛选出2 248个差异表达基因,其中553个基因表达上调,1 695个基因表达下调(P <0.05);GO功能注释分析和KEGG通路的富集分析结果显示,参与T细胞免疫功能相关的生物学过程,如TCR信号通路、T细胞增殖和活化等的差异表达基因发生显著富集;TCR信号通路,T细胞增殖、活化和凋亡通路中促进T细胞增殖活化的部分核心基因表达明显上调,而抑制T细胞活化的部分核心基因表达则明显下调。结论:上调TCRζ后CML患者T细胞的活化和增殖能力明显改善,其分子机制可能与促进T细胞活化增殖的相关基因明显上调,抑制T细胞活化和促进细胞凋亡相关基因表达的下调有关。Objective:To analyze the changes in the gene expression profile of T cells in CML patients after TCRζup-regulation expression,and to explore the molecular mechanism of T cell reactivation after transgenic up-regulation of TCRζ.Methods:The peripheral blood mononuclear cells(PBMCs) from 3 newly untreated chronic-stage CML patients were collected,and the CD3+T cells were obtained by MACS method.The TCRζ-IRES2-EGFP(experimental group)and pIRES2-EGFP(control group) plasmids were transfected into T cells by nuclear transfection technique.The gene expression profiles of CML T cells up-regulated TCRζ chain and control cells were detected by Affymetrix GeneChip Human Gene 2.0 ST Array.The differentially expressed genes were analyzed by GO functional annotation analysis and KEGG pathway enrichment analysis.Results:A total of 2248 differentially-expressed genes were obtained,including553 up-regulated genes and 1695 down-regulated genes in experimental group as compared with those in control group(P<0.05).The GO and KEGG enrichment analyses showed that differentially expressed genes involved in the biological processes related to T cell immune function,such as TCR signaling pathway,T cell proliferation and activation.Some of core genes involved in promoting the TCR signaling pathway,T cell proliferation,activation and apoptosis pathways were significantly up-regulated,while some core genes involved in inhibiting T cell activation were significantly downregulated.Conclusion:The molecular mechanism of the significantly improved T cell activation and proliferation ability in CML patients after TCRζ up-regulation may be related to the differential transcripts mediated signaling pathways of T cell activation,proliferation and apoptosis.

关 键 词：T细胞受体 慢性粒细胞白血病 CD3+T细胞 基因表达 

分 类 号：R733.72[医药卫生—肿瘤]