LAIR-1对GM-CSF介导的JAK/STAT信号通路的调节作用及其对Mo7e细胞功能的影响  

Regulation of LAIR-1 on JAK/STAT signaling Pathway Mediated by GM-CSF and its Effect on Biological Function of Mo7e Cell Line

作　　者：张娅薇 吴肖婕 杨蕊 樊翠 周嘉迪 薛江楠[1] ZHANG Yawei;WU Xiaojie;YANG Rui(Department of Immunology,Binzhou Medical University,Shandong 264003,China)

机构地区：[1]滨州医学院免疫学教研室,264003 [2]滨州市人民医院中心实验室,256610 [3]临沂市中医医院超声医学科,276000 [4]陕西中医药大学附属医院检验科,咸阳712000

出　　处：《医学研究杂志》2023年第11期39-44,共6页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81273200)。

摘　　要：目的研究人白细胞相关免疫球蛋白样受体1(leukocyte-associated immunoglobulin-like receptor-1,LAIR-1)对粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony-stimulating factor,GM-CSF)依赖的Mo7e细胞增殖、凋亡和迁移的影响,并探讨LAIR-1调节GM-CSF功能的分子机制,为进一步研究LAIR-1在造血细胞分化中的作用提供资料。方法采用LAIR-1慢病毒表达载体(LV-LAIR-1)转染GM-CSF依赖的Mo7e细胞,建立稳定高表达LAIR-1的Mo7e-LAIR-1细胞株,并采用流式细胞术、Western blot法检测、反转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)以及激光扫描共聚焦显微镜(laser scanning confocal microscopy,LSCM)鉴定LAIR-1分子的表达;采用CCK-8试剂盒、FITC Annexin V凋亡试剂盒以及Transwell实验分别检测LAIR-1对Mo7e细胞功能的影响;采用Western blot法检测LAIR-1对酪氨酸蛋白激酶/信号转导及转录激活因子(janus kinase/signal transducer and activator of transcription,JAK/STAT)信号通路影响。结果经LV-LAIR-1转染的Mo7e细胞高表达LAIR-1分子;与对照组比较,LAIR-1能够显著抑制Mo7e细胞的增殖(P<0.001),促进细胞迁移(P<0.05),但对细胞凋亡无明显影响(P>0.05);GM-CSF能够上调JAK2、STAT1、STAT3和STAT5的磷酸化水平,而LAIR-1的表达能够明显抑制STAT1、STAT3和STAT5蛋白酪氨酸的磷酸化水平。结论免疫抑制性受体LAIR-1可能通过抑制细胞因子GM-CSF介导的JAK/STAT信号通路的活化,参与对髓系造血细胞分化的调节。Objective To investigate the mechanism of leukocyte-associated immunoglobulin-like receptor-1(LAIR-1)in regulation of the granulocyte-macrophage colony-stimulating factor(GM-CSF)mediated janus kinase/signal transducer and activator of transcription(JAK/STAT)signaling pathway transduction and its influence on the function of Mo7e cell line,and to provide information for further studies on the role of LAIR-1 in hematopoietic cell differentiation.Methods Lentiviral vector of LAIR-1(LV-LAIR-1)was transferred into GM-CSF-dependent Mo7e cells to establish stable and high expression of LAVI-1 Mo7e-LAVI-1 cell line,and the LAIR-1 expression was measured by flow cytometry assay,Western blot,reverse transcription polymerase chain reaction(RT-PCR)and laser scanning confocal microscopy(LSCM).The influence of LAIR-1 on the function of Mo7e cells was detected by the CCK-8 kit,FITC Annexin V apoptosis kit and Transwell experiments.The effect of LAIR-1 on JAK/STAT signaling pathway was measured by Western blot.Results LV-LAIR-1 was highly expressed in Mo7e cells transfected by LV-LAIL-1.Compared with control group,LAIR-1 could significantly inhibit the proliferation of Mo7e cells(P<0.001),and promoted the migration of Mo7e cells,but has no significant effect on cell apoptosis.GM-CSF can upregulate the phosphorylation levels of JAK2,STAT1,STAT3 and STAT5,while the expression of LAIR-1 can significantly inhibit the phosphorylation levels of STAT1,STAT3 and STAT5 proteins.Conclusion LAIR-1 as an immune inhibitory receptor can inhabit GM-CSF-mediated JAK-STAT signaling pathway and may play a role in myeloid cell development.

关 键 词：LAIR-1 GM-CSF 酪氨酸磷酸化 增殖 迁移 凋亡 

分 类 号：R392.11[医药卫生—免疫学]