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巨噬细胞集落刺激因子受体在人白血病细胞系中的表达、特性及作用

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

作者信息

Tang Shengsong, Chen Guibin, Rao Qing, Geng Yiqi, Wu Kefu

机构信息

State Key Laboratory of Experimental Hematology, Institute Of Hematology, Peking Union Medical College and Chinese Academy of Medical Sciences, Tianjin 300020, China.

出版信息

Zhonghua Bing Li Xue Za Zhi. 2002 Jun;31(3):240-4.

Abstract

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.

摘要

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、特性及作用。

方法

采用ABC免疫过氧化物酶法、间接免疫荧光染色、流式细胞术及蛋白质印迹法,对3名健康人外周血单个核细胞(PBMCs)、5名健康人脐血单个核细胞(CBMCs)以及4种人骨髓单核细胞白血病细胞系J6-1、J6-2、K562和HL-60进行研究。

结果

在4种人白血病细胞系中,M-CSF-R定位于细胞质、细胞核及细胞膜;未受PHA刺激的正常人PBMCs未检测到M-CSF-R表达。受PHA刺激的人PBMCs表达低水平的M-CSF-R。J6-1、J6-2、K562和HL-60细胞膜结合型M-CSF-R(M-CSF-mR)表达频率分别为78.9%、72.6%、54.9%和58.0%。细胞质及细胞核相关型M-CSF-R(M-CSF-cnR)频率分别为52.3%、44.3%、28.0%和65.3%。在HL-60细胞的细胞质和细胞核中均检测到一种分子量为120 000的M-CSF-R形式。上述白血病细胞中M-CSF-cnR的半衰期长于受刺激CBMCs中相应M-CSF-R的半衰期,除K562细胞中的M-CSF-mR外,白血病细胞中M-CSF-mR的半衰期延长。抗M-CSF-R单克隆抗体和重组人M-CSF可溶性受体均可使HL-60细胞生长停滞于G(0)/G(1)期,并抑制HL-60细胞在软琼脂中形成集落。

结论

M-CSF-R在白血病细胞中的表达具有异质性。细胞内M-CSF-R的蓄积导致白血病细胞中细胞内M-CSF-R降解率降低,这可能是一种潜在的有丝分裂信号。M-CSF-R介导的信号对HL-60细胞的生长至关重要且必不可少。

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