【摘要】 目的:探讨脐血间充质干细胞对急性髓细胞白血病细胞株HL-60、急性淋巴细胞白血病细胞株Jurkat的增殖和凋亡的影响以及CXCL12/CXCR4信号轴的作用。方法:将HL-60细胞株、Jurkat细胞株分别与人脐血间充质干细胞（HUCMSC）建立共培养模型,并加用G-CSF、AMD3100单药或联合处理共培养模型。采用CCK-8法检测细胞活力,用Annexin-V/PI试剂盒检测细胞凋亡和细胞周期,流式细胞术和蛋白印迹法分别检测白血病细胞膜表面CXCR4蛋白及总CXCR4蛋白表达情况。结果:与HUCMSC共培养后HL-60细胞和Jurkat细胞增殖活力降低,凋亡减少,二者G₀/G₁期细胞增多;而加入G-CSF和AMD3100可进一步降低与HUCMSC共培养体系中的HL-60和Jurkat的细胞活力,破坏HUCMSC对HL-60和Jurkat细胞的抑凋亡作用,使2种细胞G₀/G₁期细胞比例减少,且2药联合时作用更明显。G-CSF可同时降低白血病细胞膜表面CXCR4蛋白和细胞质内总CXCR4蛋白的表达,而AMD3100只能降低白血病细胞膜表面CXCR4表达,对细胞质内总CXCR4蛋白表达无影响。结论:HUCMSC可抑制急性白血病细胞增殖及凋亡,使G₀/G₁期细胞增多;而AMD3100通过降低白血病细胞膜的CXCR4表达、G-CSF通过同时降低胞质总CXCR4蛋白和胞膜CXCR4蛋白表达来阻断CXCL12/CXCR4信号轴,减弱白血病细胞与微环境之间的联系,在HUCMSC抑制白血病细胞生长增殖、促进凋亡的基础上,进一步减弱其细胞活力,促进其凋亡,降低白血病细胞G₀/G₁期细胞比例,CXCL12/CXCR4信号轴在HUCMSC抑制白血病细胞凋亡中起重要作用。更多还原

【Abstract】 Objective: To investigate the effects of human umbilical cord blood-derived mesenchymal stem cells（ HUCMSC） on the leukemic cell line HL-60 and acute lymphoblastic leukemia cell line Jurkat as well as the role of CXCL12/CXCR4. Methods: HL-60 cells and Jurkat cells were co-cultured with human umbilical cord blood mesenchymal stem cell（ HUCMSC）,and the model was treated with G-CSF,AMD3100 and their combination. The cell viability and cell cycle were measured by Cell Counting Kit-8（ CCK-8）,the apoptosis and the cell-cycle analysis were assessed by flow cytometry with the Annexin V/PI double staining. The expression of surface CXCR4 protein and total CXCR4 protein of leukemic cells were detected by flow cytometry and Western blot respectively. Results: HUCMSC could decrease the viability of HL-60 cells and Jurkat cells,as well as the percentage of apoptotic cells,they could also increase the number of G₀/G₁ cells,while G-CSF and AMD3100 could reduce the proliferation of HL-60 cells and Jurkat cells in HUCMSC co-culture model,destructed the anti-apoptotic effect of HUCMSC on HL-60 cells and Jurkat cells,and the combination of 2 drugs resulted in a synergistic effect. The G-CSF could reduce the expression of surface CXCR4 protein and total CXCR4 protein in leukemic cells,while AMD3100 could only decrease the expression of surface CXCR4 protein of leukemia cell membrane,having no effect on the expression of CXCR4 protein in cytoplasm. Conclusion: Human umbilical cord blood mesenchymal stem cells can inhibit the proliferation and apoptosis of acute leukemia cells and increase the number of G₀/G₁ phase cells in leukemic cells. The AMD3100 can decrease the expression of surface CXCR4 protein in leukemia cells,G-CSF can decrease expression of total CXCR4 protein as well as membrane CXCR4 protein.Both of them can block the CXCL12/CXCR4 signal axis,weakening the relationship between leukemia cells and microenvironment.And on the basic of HUCMSC influenced leukemia cells’ growth and proliferation,the cell viability will be weakened,its apoptosis will be promoted,and the percentage of G₀/G₁ phase cells in leukemia cells will be decreased.更多还原