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目的探索骨髓基质细胞增强白血病细胞抗凋亡、抗药特性的可能机制。方法应用Percoll分离骨髓单个核细胞体外培养骨髓基质细胞模拟骨髓微环境功能,与白血病细胞株Jurkat细胞体外共培养。0.5μmol/LDNR处理Jurkat细胞诱导凋亡,应用AnnexinV/PI双标法流式细胞仪检测白血病细胞凋亡率。PI染色流式细胞仪检测细胞周期分布。结果0.1～2.0μmol/L浓度范围DNR作用一定时间后,Jurkat细胞发生的凋亡率随药物浓度的增加与作用时间的延长而升高。共培养后骨髓基质细胞抑制药物诱导的白血病细胞凋亡,与单独悬浮培养组比较白血病细胞凋亡率有显著降低(8.39±4.08)%比(16.02±1.00)%,P<0.05。白血病骨髓基质对白血病细胞的屏蔽效应强于正常骨髓基质细胞(白血病细胞凋亡率分别是(5.73±1.78)%比(8.39±4.08)%,P<0.05。DNR处理共培养组Jurkat细胞G0/G1期比例高于悬浮培养DNR处理组,而正常与白血病骨髓基质细胞屏蔽的白血病细胞G0/G1期阻滞现象无显著性差异(47.96±5.88)%比(39.25±3.04)%,P>0.05。结论一定浓度的DNR在体外可诱导Jurkat细胞凋亡。骨髓基质细胞抑制化疗药物诱导的白血病细胞凋亡,提示骨髓微环境在骨髓白血病细胞获得耐药、抗凋亡特性以及残留白血病形成过程中起着重要促进作用。细胞周期分布实验结果显示骨
Objective To explore the possible mechanism of bone marrow stromal cells enhancing anti-apoptosis and drug resistance of leukemia cells. Methods Percoll was used to separate bone marrow mononuclear cells in vitro to culture bone marrow stromal cells to simulate the function of bone marrow microenvironment and co-culture with leukemia cell line Jurkat cells in vitro. The apoptosis of Jurkat cells was induced by 0.5μmol / L LDNR, and the apoptosis rate of leukemic cells was detected by Annexin V / PI double-labeled flow cytometry. PI staining flow cytometry to detect cell cycle distribution. Results The apoptosis rate of Jurkat cells increased with the increase of drug concentration and the prolongation of action time after DNR of 0.1 ~ 2.0μmol / L for a certain period of time. Bone marrow stromal cells inhibited the drug-induced apoptosis of leukemia cells after co-culture. The apoptosis rate of leukemia cells was significantly lower than that of the suspension culture alone group (8.39 ± 4.08)% (16.02 ± 1.00)%, P <0.05. The leukemic bone marrow stromal cells had stronger shielding effect on leukemic cells than that of normal bone marrow stromal cells (5.73 ± 1.78% vs 8.39 ± 4.08%, P <0.05, respectively) .DNR treatment co-cultured Jurkat cells G0 / G1 phase ratio was higher than that of suspension culture DNR group, while there was no significant difference (47.96 ± 5.88)% (39.25 ± 3.04)%, P> 0.05 between normal and leukemia bone marrow stromal cells leukemia cells G0 / G1 phase arrest .Conclusion A certain concentration of DNR can induce apoptosis of Jurkat cells in vitro.Bone marrow stromal cells inhibit chemotherapeutic drug-induced apoptosis of leukemia cells, suggesting that the bone marrow microenvironment in the myeloid leukemia cells get drug resistance, anti-apoptotic characteristics and residual leukemia in the process Plays an important role in promoting the cell cycle distribution experimental results show bone