阻抑正常骨髓基质SDF-1作用对HL-60细胞生物学性质的影响

Effects of inhibition of SDF-1 activity in normal stromal cells on biological properties of human acute myelocytic leukemia cell line HL-60

目的 本研究通过应用抗CXCR4 单克隆抗体 12G5抑制趋化因子SDF 1的生物活性 ,观察HL 60细胞生物学性质的变化 ,探讨SDF 1在维持HL 60细胞生存、增殖中的作用。方法 培养HL 60细胞并与正常骨髓基质细胞共培养 ,采用 12G5阻断SDF 1生物作用 ,孵育 2h后观察对照组及单抗组HL 60细胞在骨髓基质层上的粘附情况 ,2 4h后测定细胞粘附率 ;2、4、6、8h应用台盼蓝排染法测定细胞存活率 ,MTT法检测HL 60细胞活力 ,流式细胞术观察HL 60细胞增殖周期变化。结果 ① 12G5孵育后 2 4h ,骨髓基质对HL 60的粘附率为 (19 1± 8 6) % ,显著低于对照组。②单抗孵育后HL 60细胞存活率下降 ,且随单抗孵育时间延长而逐渐下降。③细胞活性下降。④ 12G5孵育 2、6h后 ,处于增殖周期中G0 G1 期的细胞分别为 (5 4 7± 6 3 7) % ,(5 5 1± 7 0 4) % ,而S期的细胞分别为 (3 8 4± 4 5 1) % ,(3 7 1± 4 0 8) %。结论 12G5可改变HL 60细胞的生物学特性 ,从而在一定程度上抑制白血病细胞的增殖 ,影响细胞生存。

Objective To investigate the roles of chemokine SDF-1 in the maintenance of proliferation and survival of HL-60 cells and to observe the biological changes of HL-60 cells after SDF-1 activity was blocked by the monoclonal antibody (MoAb) of its receptor CXCR 4 (12G5). Methods HL-60 cells were cultured and co-cultured with normal marrow stromal cells. At 2 h after 12G5 incubation, the adhesiveness state was compared with that observed in the control group, and then cell adhesion rates were assayed in both groups at 24 h. At 2, 4, 6, and 8 h after incubation with 12G5, HL-60 cell survive rates and activities were detected by typan blue exclusion and MTT, respectively, while the cell cycles were observed by flow cytometry. Results ① Cell adhesion rate at 24 h was (19.1±8.6)% in the experiment group, significantly lower than that in the control group. ② Survive rate of leukemia cells decreased in a time-dependent manner. ③ Cell activity decreased, too. ④ HL-60 cells at G0/G1 phase at 2 and 6 h after 12G5 incubation were (54.7±6.37)% and (55.1±7.04)%, respectively, and cells at S phase were (38.4±4.51)% and (37.1±4.08)%, respectively. Conclusion 12G5 can change the biological properties of HL-60 cells and inhibit proliferation and survival of leukemia cells at a certain level.