异型巨噬细胞集落刺激因子在人白血病细胞系中的表达及性质

目的 探讨异型巨噬细胞集落刺激因子（Ｍ－ＣＳＦ）在人白血病细胞系中的表达和性质。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,蛋白免疫印迹和反向酶联ＤＮＡ蛋白质相互作用分析技术研究了４株人白血病细胞系（Ｊ６－１、Ｊ６－２、Ｋ５６２、ＨＬ６０）和正常人外周血单个核细胞Ｍ－ＣＳＦ的分布、表达及其分子大小。结果 正常人外笛血单个核细胞示见Ｍ－ＣＳＦ的表达,经植物血浆素刺激后有低水平的表达

The expression and effects of isoforms of macrophage colony stimulating factor in human leukemic cell lines

Objective To explore the expression and effects of isoforms of macrophage colonystimulating factor (MCSF) in human leukemic cell lines. Methods Three normal human peripheral blood mononuclear cells (PBMCs) and 4 human myelomonocytic leukemic cell lines including J61, J62, K562 and HL60 were studied using ABC immunoperoxidase assay, indirect immunofluorescene staining, flow cytometry, Western blot and reverse enzymelinked DNAprotein interaction assay (reverse ELDIA). Results MCSF was noticed to be localized in the cytoplasm, nucleus and at the cell membrane in 4 human leukemic cell lines; expression of MCSF was not detected in normal human PBMCs without PHA stimulation. Human PBMCs stimulated by PHA expressed a low level of MCSF. Frequencies of membrane bound MCSF expression in J61,J62,K562 and HL 60 were 71.6%,69.7%, 42.7% and 57.4% respectively. Frequencies of cytoplasm and nucleus associated MCSF were 65.7%, 45.4%, 36.5% and 72.5% respectively. The cytosolic bound MCSF was expressed in J61 cell as four isoforms with a molecular weight of 14 000, 16 000, 20 000 and 44 000. While nucleus associated MCSF expressed as two isoforms with a molecular weight of 16 000 and 20 000. AntiMCSF monoclonal antibody could dramatically inhibit proliferation of leukemic cells and its inhibitory effect was related to the levels of membrane bound MCSF expression in leukemic cells. Reverse ELDIA showed that MCSF could bind with DNA in vitro. Conclusions Expression of MCSF isoforms is heterogeneous and polymorphous in leukemic cells. Membrane bound MCSF is crucial for the proliferation of leukemic cells, which might be a DNAbound protein and could be involved in the transformation and tumorigenesis of hematopoietic cells.