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 An-nexin 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 informa-tion 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 reac-tion(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.