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背景与目的:近年研究发现,骨髓基质细胞分泌的趋化因子SDF-1通过其分布于白血病细胞膜表面的生理性受体CXCR4可能参与了白血病细胞的髓内屏障,然而,SDF-1/CXCR4系统与白血病细胞之间的相互关系尚不清楚,因此,本研究采用抗CXCR4单克隆抗体12G5阻抑SDF-1活性,观察与白血病骨髓基质细胞共培养的HL-60细胞粘附性及细胞增殖的变化,以探讨从阻抑SDF-1活性入手治疗残留白血病的可能性。方法:培养并共培养HL-60细胞,采用12G5(10μg/ml)阻断SDF-1生物作用,观察HL-60细胞在骨髓基质层上的粘附情况并测定细胞粘附率,通过流式细胞仪检测凋亡率,并观察其增殖周期的变化;应用台盼蓝拒染法检测细胞存活情况,绘制细胞生长曲线。结果:(1)12G5孵育后24h,骨髓基质对HL-60的粘附率为(39.4±7.9)%,对照组为(51.4±5.9)%,二者具有显著差异(P<0.05)。(2)比较细胞增殖周期及凋亡率,实验组中G0/G1期细胞比例为(55.2±4.9)%,S期为(30.4±4.1)%,G2/M期为(14.4±5.2)%,细胞凋亡比例为(9.0±1.7)%;对照组中G0/G1期细胞比例为(44.7±2.2)%,S期为(45.3±3.7)%,G2/M期为(10.0±2.6)%,细胞凋亡比例为(4.0±2.4)%。(3)12G5孵育48h后,HL-60细胞生存率明显下降,增殖减缓。结论:12G5能在一定程度上抑制HL-60细胞的粘附性及增殖活性,因此,通过1
BACKGROUND & OBJECTIVE: In recent years, it has been found that the chemokine SDF-1 secreted by bone marrow stromal cells may be involved in the intramedullary barrier of leukemia cells through its physiological receptor CXCR4 distributed on the membrane of leukemia cells. However, SDF-1 / CXCR4 system And leukemia cells is not yet clear, therefore, in this study, anti-CXCR4 monoclonal antibody 12G5 SDF-1 activity was inhibited to observe the adhesion and cell proliferation HL-60 cells co-cultured with leukemia bone marrow stromal cells Changes in order to explore the inhibition of SDF-1 activity to start the possibility of residual leukemia. Methods: HL-60 cells were cultured and co-cultured. The biological effect of SDF-1 was blocked by 12G5 (10μg / ml). The adhesion of HL-60 cells to the stromal layer of bone marrow was observed and the cell adhesion rate was determined. Apoptosis rate was detected by cytometry and the change of proliferation cycle was observed. Cell viability was measured by trypan blue exclusion method and cell growth curve was drawn. Results: (1) Adhesion rate of HL-60 to bone marrow stroma was (39.4 ± 7.9)% at 12G5 and (51.4 ± 5.9)% at 12 h, respectively. There was significant difference between the two groups (P <0.05). (2) Compare the cell proliferation cycle and apoptosis rate, the percentage of cells in G0 / G1 phase was (55.2 ± 4.9)%, the S phase was (30.4 ± 4.1)%, the G2 / M phase was (14.4 ± 5.2) (9.0 ± 1.7)%. The percentage of cells in G0 / G1 phase was (44.7 ± 2.2)% in control group, (45.3 ± 3.7)% in S phase and (10.0 ± 2.6) in G2 / %, The rate of apoptosis was (4.0 ± 2.4)%. (3) After 48G incubation for 12G5, the survival rate of HL-60 cells decreased significantly and the proliferation slowed down. Conclusion: 12G5 can inhibit the adhesion and proliferation activity of HL-60 cells to a certain extent. Therefore,