摘要
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 study the effect of thalidomide and arsenic trioxide on the proliferation and apoptosis in human myelodysplastic syndrome cell line MUTZ-1 and explore its possible mechanism. Method： MUTZ-1 cells were cultured with different concentration of thalidomide alone, arsenic trioxide alone, and thalidomide plus arsenic trioxide for 48 h. Cell proliferation was analyzed by CCK-8 test. Cell apoptosis was analyzed by flow cytometry. VEGF expression was analyzed by ELISA and semi-quantitative RT-PCR. Result： Thalidomide alone had no significant growth inhibition in MUTZ-1 cells （ P 〉 0.05）. Arsenic trioxide alone obviously inhibited cell proliferation （ P 〈0.05）. Thalidomide plus arsenic trioxide group had more inhibition effect, which revealed that two drugs had synergism effect in inhibiting the MUTZ-1 cells. The apoptosis rate of MUT-1 cell had no significant increase while the thalidomide concentration increased alone （r= 0.313, P 〉 0.05）. Apoptosis rate increased while Arsenic trioxide increased alone in a close dependent manner （r=0. 627, P d0.05）, and also increased in the combined group （ P 〈0. 001）. Thalidomide alone in the concentration of 10, 25 mg/L had no impaction on VEGF secretion （ P 〉 0.05）, but inhibited the VEGF expression at the concentration of 50, 100, 200 mg/L（ P 〈0. 05）, and the inhibition effect was most obviously at the 50 mg/L concentration. Arsenic trioxide alone inhibited VEGF expression in a dose dependent manner （ P 〈0.05）, but thalidomide 200 mg/L plus arsenic trioxide 10 μmol/L group had a slightly declined VEGF expression compared with thalidomide 100 mg/L plus arsenic trioxide5 μmol/L group. Conclusion：Thalidomide had no significant proliferation inhibition effect on MUTZ-1 cells. Arsenic trioxide could inhibit MUTZ-1 cells alone. And the combined group had more significantly effect than that of arsenic trioxide or thalidomide alone. The low concentration thalidomide had no effect on VEGF expression. But high concentration had an obvious inhibition effect on VEGF expression. Arsenic trioxide alone can inhibit the expression of VEGF to a certain concentration extent, in a dose-dependent manner. The combined group had more obviously inhibition effect than thalidomide or arsenic trioxide alone. Apoptosis rate of MUTZ-1 cell had no significant increase when thalidomide concentration increased alone. But the apoptosis rate increased as arsenic trioxide alone increased in a dose-dependent manner, and the combined group also had this effect.
出处
《临床血液学杂志》
CAS
2008年第6期578-582,共5页
Journal of Clinical Hematology
基金
上海市卫生局青年科研项目(No:2006Y71)
关键词
骨髓增生异常综合征
细胞系
凋亡
增殖
血管内皮生长因子
三氧化二砷
沙立度胺
Myelodysplastic syndrome
Cell lines
Apoptosis
Proliferation
Vascular endothelial growthfactor
Arsenic trioxide
Thalidomide
