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为了观察正常人骨髓成纤维样细胞系 (HFCL)对白血病细胞系HL 60细胞增殖和血管内皮生长因子(VEGF)表达的影响 ,用免疫组织化学对HFCL细胞进行组织学鉴定 ;建立HL 60细胞和HFCL细胞共培养体系 ,台盼蓝拒染法测定生长曲线 ;HE染色和瑞 姬姆萨染色后进行分裂指数 (MI)测定 ;流式细胞术检测细胞周期的变化 ;Westernblot检测增殖细胞核抗原 (PCNA)表达 ;ELISA双抗体夹心法检测HL 60细胞表达VEGF水平。结果发现 ,与HFCL细胞共培养后HL 60细胞生长受抑 ,且与HFCL细胞直接接触组的抑制作用大于用跨室共培养组 ,其分裂指数表现为单独HL 60细胞组大于用跨室共培养组 ,更大于与HFCL细胞直接接触组。同时发现HL 60细胞与HFCL细胞共培养后G1期细胞增多 ,S期细胞减少 ,且PCNA表达下调 ,以直接接触组为甚。HL 60细胞与HFCL细胞共培养后 ,培养上清的VEGF水平减低 ,直接接触组和跨室共培养组下降程度一样 ,而与HL 60细胞共培养 ,HFCL细胞增殖无明显变化。结论 :正常人骨髓成纤维样细胞能抑制白血病细胞系HL 60的增殖 ,抑制细胞周期 ,降低VEGF因子的表达
To observe the effect of normal human bone marrow fibroblastoid cell line (HFCL) on HL-60 cell proliferation and vascular endothelial growth factor (VEGF) expression, HFCL cells were identified by immunohistochemistry. HL 60 cells and HFCL cell co-culture system, and the growth curve was determined by trypan blue exclusion method. The mitotic index (MI) was determined by HE staining and Ringer’s method. The cell cycle was detected by flow cytometry. The proliferating cell nuclear antigen (PCNA) ). The level of VEGF was detected by double antibody sandwich ELISA in HL 60 cells. The results showed that the inhibition of HL-60 cells co-cultured with HFCL cells was inhibited, and the direct contact with HFCL cells was greater than the inhibition effect of intercross culture group, the division index showed HL60 cell group than co-culture Group, more in direct contact with HFCL cells. At the same time, it was found that the number of cells in G1 phase and the number of S phase in HL 60 cells co-cultured with HFCL cells were decreased, and the expression of PCNA was down-regulated in direct contact group. After co-cultured HL-60 cells with HFCL cells, the level of VEGF in the culture supernatant decreased. The direct contact group and transwell co-culture group decreased in the same extent. However, the co-culture with HL-60 cells showed no significant changes in the proliferation of HFCL cells. Conclusion: Normal human bone marrow fibroblast-like cells can inhibit the proliferation of leukemia cell line HL 60, inhibit the cell cycle and decrease the expression of VEGF