抗CXCR_4单克隆抗体对与正常骨髓基质细胞共培养的HL-60细胞Bcl-2、PCNA、Fas蛋白表达的影响

Effects of anti- CXCR4 monoclonal antibody on the expression of Bcl-2, PCNA and Fas proteins of human acute myelocytic leukemia cell line HL-60 co-cultured with normal bone marrow stromal cells

目的 目前研究发现 ,骨髓基质细胞分泌的基质衍生因子 1(stromalcellderivedfactor 1,SDF 1)及其受体CXCR4可能参与髓内残留病变的形成。因此 ,本研究着重探讨了阻抑SDF 1活性对与正常骨髓基质细胞共培养的HL 60细胞增殖、生存的影响。方法 培养并共培养HL 60细胞。实验分为两组 ,实验组采用抗CXCR4单克隆抗体 12G5( 2 0ug/ml)阻断SDF 1生物作用 ,孵育 2 4小时后采用免疫组化染色检测HL 60细胞Bcl 2、PCNA、Fas蛋白表达。 结果 免疫组化染色显示 ,实验组中Bcl 2、PCNA及Fas阳性率分别为 ( 15 .7± 4.9) %、( 4 2 .1± 3 .9) %及 ( 3 9.7± 7 5 ) % ,而对照组分别为 ( 3 1.6± 5 .2 ) %、( 67.4± 8.5 ) %和 ( 2 4.1± 6.7) % ,t检验显示 12G5下调HL 60细胞Bcl 2及PCNA蛋白表达 ,上调Fas蛋白表达。结论 12G5可在一定程度抑制HL 60细胞增殖 ,促进凋亡 ,影响其生存。

Objective It has been shown recently that stromal cell derived factor-1 (SDF-1), secreted by bone marrow stromal cells, and its receptor CXCR4 are concerned with MRD. We investigate the effects of chemokine SDF-1 on proliferation and survival of acute myelocytic leukemia cells HL-60 co-cultured with normal bone marrow stromal cells. Methods HL-60 cells were cultured and co-cultured with normal marrow stromal cells. All samples were divided into two groups, experiment and control. In experiment group, SDF-1 activity was blocked by anti-CXCR4 monoclonal antibody 12G5 (20ug/ml). By 24 h after incubation, the expression of Bcl-2, PCNA, and Fas proteins were detected by the immunohistochemical technique.Results In experiment group, the positive rate of Bcl-2 was 15.7%±4.9%; PCNA, 42.1%±3.9%; and Fas, 39.7%±7.5%. In control group however, the indexes were 31.6%±5.2%, 67.4%±8.5% and 24.1%±6.7%, respectively. The results revealed that 12G5 down-regulated the expression of Bcl-2 and PCNA, but up-regulated the expression of Fas protein. Conclusion 12G5 can inhibit proliferation of HL-60 cells and increase cell apoptosis, and accordingly influence the survival of leukemia cells.