人成纤维样基质细胞系对HL-60细胞增殖和VEGF表达的影响

为了观察正常人骨髓成纤雏样细胞系(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，台盼蓝柜柒法测定生长曲线；既染色和瑞-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；Western blot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。

Effects of Human Fibroblastoid Stromal Cell Line on Proliferation of HL-60 Cells and Expression of VEGF

To investegate the effects of normal human bone marrow fibroblastoid s tromal cell line (HFCL) on the proliferation of acute myeloid leukemia cell line HL-60 and expression of vascular endothelial growth factor (VEGF), establishing cocult ure system of leukemia cell line HL- 60 and HFCL, growth data was o btained by cell counting. Mitotic index (MI) was observed under Wright-Giemsa staining. Flow cytometry and Western blot were used as assays for cell cycle and expression of proliferating cell nuclear antigen (PCNA) separately. VE GF levels were evaluated by using commercial ELISA kits. The results showed that compared with HL-60 cel ls without HFCL cells, the proliferation of HL-60 cells in direct contact with HFCL cells and with HFCL cells separated by transwell was inhibited.The MI of HL -60 cells without HFCL cells was highest followed by HL-60 cells separated by tra nswell and HL-60 cells in direct contact with HFCL cells. The expression of PCN A in HL-60 cells with HFCL cells were lower than HL-60 cells without HFCL cell s. Meanwhile, the percentage of HL-60 cells in G 1 phase cocultured with HF CL cells was higher than that without HFCL cells while the percentage of S phase ce lls was lower. The levels of VEGF in HL-60 cells with HFCL cells were lower tha n that in HL-60 cells alone. In conclusion, the normal bone marrow fibroblastoi d stromal cells inhibited the proliferation of HL-60 cells as well as the expre ssion of VEGF.