巨噬细胞集落刺激因子受体在白血病细胞系中的表达和作用

The expression, characterization and roles of macrophage colony-stimulating factor receptor in human leukemia cell lines

目的 探讨巨噬细胞集落刺激因子受体 (M CSF R)在人白血病细胞系中的表达及其作用。方法 采用ABC免疫酶标技术、间接免疫荧光染色、流式细胞计数和蛋白免疫印迹研究了 4株人白血病细胞系 (J6 1、J6 2来源于人粒 单型白血病 ,HL 6 0为来源于人急性粒细胞白血病的髓样细胞系 ,K5 6 2为来源于人慢性白血病急变期胸水的髓样细胞系 )和正常人外周血单个核细胞及正常人脐带血单个核细胞M CSF R的分布、表达、分子大小及其作用。结果 正常人外周血单个核细胞未见M CSF R的表达 ,经PHA刺激后有低水平的表达 ;4株人白血病细胞系都高表达M CSF R ,分布于细胞膜、细胞质和细胞核 ;J6 1、J6 2、K5 6 2和HL 6 0细胞质和胞核M CSF R(M CSF cnR)平均阳性率分别为5 2 3%、4 4 3%、2 8 0 %、6 5 3% ;胞膜M CSF R (M CSF mR)阳性率分别为 78 9%、72 0 %、5 4 9%、5 8 0 % ;高于正常人外周血单个核细胞的表达水平。HL 6 0细胞质和胞核中有一种相对分子质量为12 0 0 0 0的M CSF R分子 ;4种白血病细胞表达的M CSF cnR的半衰期分别高于正常人脐带血单个核细胞经PHA诱导表达的M CSF cnR的半衰期 ;J6 1、J6 2、HL 6 0细胞表达的M CSF mR的半衰期亦明显延长。抗M CSF R单抗和人重组的M CSF R可溶性受体均能使 4种?

Objective To explore the expression, characteristics and roles of macrophage colony stimulating factor receptor (M CSF R) in human leukemia cell lines Methods Peripheral blood mononuclear cells (PBMCs) collected from 3 healthy persons, cord blood mononuclear cells (CBMCs) collected from 5 healthy persons and 4 human myelomonocytic leukemia cell lines including J6 1, J6 2, K562 and HL 60 were studied by using ABC immunoperoxidaes assay, indirect immunofluorescene staining, flow cytometry, and Western blot Results M CSF R was noticed to be localized in the cytoplasm, nucleus and at the membrane in 4 human leukemia cell lines; expression of M CSF R was not detected in normal human PBMCs without PHA stimulation Human PBMCs stimulated by PHA expressed a low level of M CSF R Frequencies of membrane bound M CSF R (M CSF mR) expression in J6 1, J6 2, K562 and HL 60 were 78 9%, 72 6%, 54 9% and 58 0% respectively Frequencies of cytoplasm and nucleus associated M CSF R (M CSF cnR) were 52 3%, 44 3%, 28 0% and 65 3% respectively One form of M CSF R with a molecular weight of 120 000 was detected both in the cytoplasm and nucleus of HL 60 cells The half life of M CSF cnR in leukemia cells mentioned above was longer than that of corresponding M CSF R in stimulated CBMCs, and the half life of M CSF mR in leukemia cells was extended except that of M CSF mR in K562 cells Both anti M CSF R monoclonal antibody and recombinant human M CSF soluble receptor could cause the growth arrest of HL 60 cell in G 0/G 1 phase, and could inhibit the formation of colony of HL 60 cell in soft agarose Conclusions Expression of M CSF R in leukemia cells is heterogeneous The accumulation of cellular M CSF R results in the low degradation rate of cellular M CSF R in leukemia cells, which could be a potential mitotic signal Signal mediated by M CSF R is important and necessary for the growth of HL 60 cell