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为探讨急性髓系白血病 (AML)原代细胞自分泌的粒细胞—巨噬细胞集落刺激因子 (GM -CSF)和重组人粒细胞—巨噬细胞集落刺激因子 (rhGM -CSF)对原代AML细胞凋亡的影响 ,应用放射免疫技术检测GM -CSF水平 ,DNA电泳检查梯状条带及流式细胞仪检测细胞凋亡率和Bcl- 2蛋白表达的阳性细胞率和相对荧光强度 (MFI)。结果在 5例原代AML细胞体外培养中 ,3例的培养上清液可检测到GM -CSF ,其自发细胞凋亡率比其余 2例明显减少 ;bcl- 2蛋白表达的MFI明显增强。阿糖胞苷 (Ara -C) 5μg/ml联合rhGM -CSF 5ng/ml作用AML细胞 2 4h ,细胞凋亡率比单独Ara -C组显著增加达 (56 4± 7 4 ) % (P <0 0 5)。说明有自分泌GM -CSF的AML细胞自发凋亡率低 ,其机制可能与GM -CSF提高bcl- 2蛋白表达有关 ;rhGM -CSF能够增强Ara -C诱导AML细胞凋亡
To investigate the effect of GM-CSF and rhGM-CSF on autologous acute myeloid leukemia (AML) The level of GM-CSF was detected by radioimmunoassay. The rate of apoptosis and the expression of Bcl-2 were detected by DNA ladder and flow cytometry. The positive cell ratio and relative fluorescence intensity (MFI) . Results In 5 cases of primary AML cells cultured in vitro, GM-CSF could be detected in 3 of the culture supernatants. The rate of spontaneous apoptosis was significantly decreased compared with the other 2 cases. The MFI of bcl-2 protein expression was significantly increased. Apoptosis rate of AML cells treated with Ara-C 5μg / ml and rhGM-CSF 5ng / ml for 24 hours increased significantly (56 4 ± 7 4%) compared with Ara-C alone group (P 0 0 5). The results indicated that the autophagic apoptosis rate of AML cells with autocrine GM-CSF was low, which may be related to the increased expression of bcl-2 protein by GM-CSF. RhGM-CSF could enhance the apoptosis of AML cells induced by Ara-C