抑制性受体LAIR-1在巨核细胞系Dami上的表达及PMA对其表达的影响

Expression of the inhibitory receptor LAIR-1 in megakaryocytic cell line Dami and the influence of phorbol myristate acetate on its expression

研究LAIR-1在巨核细胞系上的表达,探讨其表达与巨核细胞分化发育的关系,为深入研究LAIR-1在巨核细胞造血中的作用奠定基础。分别通过RT-PCR技术和流式细胞术在核酸和蛋白质水平上检测LAIR-1在巨核细胞系HEL、Dami、Mo7e中的表达。应用PMA和TPO刺激Dami细胞分化,流式细胞术和激光共聚焦显微镜技术检测Dami细胞分化过程中CD41a、CD42b和LAIR-1分子的表达。结果显示:Dami细胞中LAIR-1分子在核酸水平上呈现高表达,但膜蛋白表达水平很低。在PMA诱导Dami细胞分化过程中LAIR-1分子膜表达水平显著上调,PMA未刺激的Dami细胞中LAIR-1分子的阳性表达率为2.78%±0.50%,PMA刺激6 d后,Dami细胞中LAIR-1分子的阳性表达率为49.52%±1.86%。PMA可显著诱导Dami细胞中LAIR-1分子的表达;Dami细胞的分化与LAIR-1分子的表达水平之间存在因果关系;由于PMA是PKC的激活剂,LAIR-1分子表达水平可能受PKC信号通路的调节。

To investigate the expression of the inhibitory receptor LAIR-1 on megakaryocytes,and the relationship between the expression of LAIR-1 and the development of megakaryocytes in order to lead a better insight in the function of LAIR-1 on the megakaryocytopoiesis.RT-PCR and flow cytometry were used to determine the DNA and protein contents of LAIR-1 on megakaryocytic cell lines HEL,Dami and Mo7e.After treatment with phorbol myristate acetate（PMA） and TPO,flow cytometry and laser co-focal technique were used to detect the expression of CD41a,CD42b and LAIR-1 in the differentiation of Dami cells.It was demonstrated that the expression of LAIR-1 in Dami cells was high at nuclear acid level,but low for the membrane protein.PMA could significantly induce the expression of LAIR-1 on Dami cells.The positive expression rate of LAIR-1 on non-PMA treated Dami cells was 2.78±0.50%,and the rate was 49.52±1.86% after treated with PMA for 6 days.It is evident that PMA significantly induces the expression of LAIR-1 on Dami cells and the differentiation of Dami cells has causal relationship with the expression of LAIR-1.Because PMA is the activator of PKC,the expression level of LAIR-1 may be regulated by PKC signaling pathway.