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目的:探讨沙立度胺和三氧化二砷对人类骨髓增生异常综合征细胞株的影响及作用机制。方法:采用CCK-8法检测沙立度胺、三氧化二砷、以及二者联合用药对人MUTZ-1细胞株增殖是否有抑制作用。采用ELISA法、半定量RT-PCR方法分别检测沙立度胺、三氧化二砷、以及2者联合对人类骨髓增生异常综合征细胞株MUTZ-1血管内皮生长因子(VEGF)是否有抑制作用,并用流式细胞术对细胞株在3组药物作用后行凋亡检测。结果:CCK-8法提示沙立度胺体外对MUTZ-1细胞增殖无明显抑制作用(P>0.05)。三氧化二砷对MUTZ-1细胞有明显生长抑制作用(P<0.05)。联合用药组的抑制作用明显高于三氧化二砷、沙立度胺单独用药组,两药存在协同作用。ELISA法、半定量RT-PCR方法检测显示沙立度胺组药物浓度分别为10、25mg/L的2组对MUTZ-1细胞分泌的VEGF无明显抑制作用(P>0.05)、50、100、200mg/L的3组对MUTZ-1细胞分泌的VEGF有明显抑制作用(P<0.05),其中以50mg/L为最明显,三氧化二砷组从0.05、0.25、0.5、2.5、5至10μmol/L随着浓度逐渐增高细胞株分泌VEGF逐渐下降,呈剂量依赖性分泌抑制(P<0.05)。沙立度胺凋亡率随着药物浓度增加无明显升高(r=0.313,P>0.05),三氧化二砷组的凋亡率随着药物浓度增加升高,呈剂量依赖(r=0.627,P<0.05),联合用药组随着药物浓度增加表达亦下降,呈剂量依赖(P<0.001)。结论:沙立度胺体外对MUTZ-1细胞无明显生长抑制作用。三氧化二砷对MUTZ-1细胞有明显生长抑制作用。联合用药组的抑制作用明显高于三氧化二砷、沙立度胺单独用药组。低浓度沙立度胺对VEGF表达无明显的抑制作用。高浓度沙立度胺对VEGF表达有明显的抑制作用。三氧化二砷在一定浓度范围内能抑制VEGF表达,呈浓度依赖。联合用药组的抑制作用明显高于三氧化二砷、沙立度胺单独用药组。沙立度胺凋亡率随着药物浓度增加无明显升高,三氧化二砷组的凋亡率随着药物浓度增加升高,呈剂量依赖,联合用药组随着药物浓度增加表达亦下降,呈剂量依赖。
Objective: To investigate the effect and mechanism of thalidomide and arsenic trioxide on human myelodysplastic syndrome cell line. Methods: CCK-8 method was used to detect thalidomide, arsenic trioxide, and the combination of the two on human MUTZ-1 cell proliferation was inhibited. ELISA and semi-quantitative RT-PCR methods were used to detect whether thalidomide, arsenic trioxide and their combination could inhibit the growth of human myelodysplastic syndrome cell line MUTZ-1 vascular endothelial growth factor (VEGF) Cell surgery on the cell lines in the three groups of drugs after apoptosis detection. Results: CCK-8 method showed thalidomide had no significant inhibitory effect on the proliferation of MUTZ-1 cells in vitro (P> 0.05). Arsenic trioxide significantly inhibited the growth of MUTZ-1 cells (P <0.05). The inhibitory effect of the combination group was significantly higher than that of arsenic trioxide, thalidomide alone group, the two drugs synergistic effect. ELISA and semi-quantitative RT-PCR showed that the two concentrations of 10 and 25 mg / L of thalidomide group did not significantly inhibit the VEGF secretion of MUTZ-1 cells (P> 0.05) 200mg / L of 3 groups of MUTZ-1 cells secreted VEGF was significantly inhibited (P <0.05), of which 50mg / L was the most obvious, arsenic trioxide from 0.05,0.25,0.5,2.5,5 to 10μmol / L with With the increasing of concentration, the secretion of VEGF in the cell lines decreased gradually, and the secretion of VEGF was inhibited in a dose-dependent manner (P <0.05). The apoptosis rate of thalidomide was not significantly increased with the increase of drug concentration (r = 0.313, P> 0.05). The apoptosis rate of arsenic trioxide group increased with the increase of drug concentration in a dose dependent manner (r = 0.627, P < 0.05). The combination group also decreased with the increase of drug concentration in a dose-dependent manner (P <0.001). Conclusion: Thalidomide has no significant inhibitory effect on MUTZ-1 cells in vitro. Arsenic trioxide significantly inhibited the growth of MUTZ-1 cells. The inhibitory effect of combination group was significantly higher than that of arsenic trioxide, thalidomide alone group. Low concentration thalidomide had no significant inhibitory effect on VEGF expression. High concentration of thalidomide significantly inhibited VEGF expression. Arsenic trioxide inhibits VEGF expression in a concentration-dependent manner in a concentration-dependent manner. The inhibitory effect of combination group was significantly higher than that of arsenic trioxide, thalidomide alone group. The apoptosis rate of thalidomide did not increase significantly with the increase of drug concentration. The apoptosis rate of arsenic trioxide group increased with the increase of drug concentration in a dose-dependent manner. The drug combination group decreased with the increase of drug concentration in a dose-dependent manner .