沙利度胺联合三氧化二砷对急性非淋巴细胞白血病细胞血管内皮生长因子表达的影响

目的 研究沙利度胺和三氧化二砷(As2O3)对急性非淋巴细胞白血病(ANLL)细胞分泌血管内皮生长因子(VEGF)的影响,并研究两者是否有协同作用.方法 收集23例ANLL患者的骨髓液,用Ficoll淋巴细胞分离液分离单个核细胞(MNC),于37℃、5%二氧化碳CO2饱和湿度条件下于RPMI1640培养液中传代培养.取对数生长期细胞以2×106/mL的细胞浓度加入24孔培养板上,沙利度胺以75、150和300μg/mL的浓度加入培养体系中,As2O3以12.5,25和50μmoL/L的浓度加入培养体系中,并将300μg/mL的沙利度胺和50μmol/L的As2O3联合加入培养体系中,同时设空白对照.上述各组分别培养48、72、96和120 h.用ELISA法检测上清液VEGF浓度.结果 沙利度胺和As2O3均可抑制ANLL细胞分泌VEGF,其抑制程度具有剂量-时间依赖性.两药联用的抑制作用更加明显.结论 白血病细胞能自分泌VEGF;沙利度胺和As2O3均能抑制ANLL细胞分泌VEGF;两者联合应用起协同作用.

Study on effect of combination of thalidomide and As2O3 on VEGF expression in ANLL

[Objective] To investigate the effect of thalidomide and As2O3 on the secretion of vascular endothelial growth factor (VEGF) of the cell of ANLL and study whether there is synergism between thalidomide and As2O3. [Methods] 23 patients with ANLL were analyzed.Heparinized marrow was taken, and putted into Ficoll-isopaque solution to purified MNC, cultured in the RPMI 1640 medium, placed in 5% CO2 saturation and 37℃ to proliferation. The cells were chosen in logarithm growth phase,cultural solution was added to obtain the cellular density of 2×106/mL and then the cells were put into 24-well plates. Thalidomide of 75, 150 and 300μg/mL, As2O3 of 12.5, 25 and 50 μg/mL,and thalidomide (300 μg/mL) combined with As2O3 (25 μg/mL) was added to substrate respectively, cultured for 48, 72, 96 and 120 hours, respectively. At the same time, a contrast group was set up. The concentration of VEGF in upper liquid was examined with ELISA method. [Result] Both thalidomide and A2O3 of different concentrations can inhibit ANLL cells from secreting VEGF. The inhibition was dose-time dependent. Their combination shows more obvious inhibition. [Conclusion] Leukemia cell can secrete VEGF selfly; Both Thalidomide and As2O3 can inhibit the expression of VEGF in leukemia cells;Thalidomide can enhance the function of anti-leukemia of As2O3.