抑制SDF-1活性对HL60细胞增殖影响的实验研究

Investigation on the effect of inhibiting the activity of SDF-1 on proliferation of human acute myelocytic leukemia cell line HL60

目的 通过应用SDF 1受体CXCR4单克隆抗体 (1 2G5 )观察抑制SDF 1活性对HL6 0细胞增殖活性的影响 ,探讨趋化因子SDF 1在维持HL6 0细胞生存能力中的作用。方法 培养骨髓基质细胞 ,并与HL6 0细胞共培养 ,采用SDF 1受体CXCR4单克隆抗体阻断SDF 1生物活性后 ,用MTT法检测HL6 0细胞活力 ,流式细胞术观察HL6 0细胞增殖周期变化、检测细胞膜表面CXCR4表达 ,同时检测CXCR4单克隆抗体应用前后HL6 0细胞内游离钙离子浓度等。结果 应用抗CXCR4单克隆抗体后 ,HL6 0细胞出现下列变化 :(1 )细胞膜表面CXCR4表达下调 ;(2 )增殖周期中G0 /G1 期的细胞增多 ,增殖期细胞减少 ;(3)细胞活性下降 ;(4 )细胞内游离钙离子浓度降低 ,并与单抗浓度呈正相关。结论 抑制SDF 1活性可在一定程度上抑制白血病细胞增殖 ,但对于抑制髓内残留病变的形成尚需进一步研究。

Objective To investigate the importance of chemokine SDF 1 in maintaining proliferation ability of acute myelocytic leukemia cell line HL60.Methods Co culturing marrow stromal cells and HL60 cell, and blocking chemokine SDF 1 activity with anti CXCR 4 McAb(12G5), then the activity of HL60 cells were detected with MTT, and the cell cycle and the expression of CXCR 4 of HL60 cells were observed with flow cytometry. Meanwhile, the internal free calcium ionic concentration of HL60 cells were tested before and after treated with 12G5. Results (1) The expression of CXCR 4 on HL60 cell membrane was down regulated; (2) HL60 cells in G 0/G 1 period were increased, and those in proliferate phase were decreased;(3)The activity of leukemia cells were reduced; (4)The internal free calcium ionic concentration of HL60 cell decreased after treated with 12G5, and was positively correlated with concentration of 12G5. Conclusion Blocking the activity of SDF 1 may inhibite the proliferation of leukemia cell at Certain level, but the effect on inhibiting the formation of marrow residual disease needs further study.