二烯丙基二硫对HL-60细胞血管内皮生长因子表达及分泌的影响

Effect of diallyl disulfide on the expression and secretion of VEGF in HL-60 cells

目的探讨二烯丙基二硫(DADS)对白血病细胞株 HL-60的生长抑制作用和对 HL-60细胞血管内皮生长因子(VFGF)mRNA 的表达及 VEGF 蛋白分泌的影响。方法采用 MTT 法观察DADS 对体外培养的 HL-60细胞的生长抑制;应用 ELISA 法检测药物作用前后 HL-60细胞培养上清液中 VEGF 蛋白的含量;半定量 RT-PCR 法检测药物作用前后 HL-60细胞 VEGF mRNA 表达水平。结果DADS 对 HL-60细胞具有明显的生长抑制效应,0.625,1.250和2.500μg/ml DADS 作用 HL-60细胞24 h 的细胞生长抑制率分别为(8.19±3.34)%,(16.79±2.07)%,(21.30±2.72)%;作用48 h 的细胞生长抑制率分别为(11.93±3.93)%.(22.81±2.31)%,(30.74±2.03)%;作用72 h 的细胞生长抑制率分别为(16.68±2.37)%,(28.54±3.26)%,(36.59±2.37)%,其对 HL-60细胞的生长抑制率与药物浓度有明显依赖关系(r>0.9,P<0.01),各实验组随作用时间增加抑制率明显增加(r>0.7,P<0.01);0.625,1.250,2.500 μg/ml DADS 作用 HL-60细胞48 h 和72 h 与空白对照相比均能下调HL-60细胞 VEGF mRNA 的表达及 VEGF 蛋白的分泌(P<0.01),三个不同浓度 DADS 组间差异均有统计学意义(P<0.01),其下调 HL-60细胞 VEGF mRNA 的表达及 VEGF 蛋白的分泌效果与药物浓度呈相关性(r>0.9,P<0.01)。结论 DADS 能明显抑制 HL-60细胞的增殖;DADS 可能通过抑制 HL-60细胞 VEGF mRNA 的表达及 VEGF 蛋白分泌发挥抗白血病的效应。

Objective To investigate the proliferation inhibition of human leukemic cell line HL-60 and the expression of vascular endothelial growth factor （VEGF） mRNA and secretion of VEGF protein in HL-60 cells treated with diallyl disulfide （DADS）. Methods MTT was used to test the cell growth, semiquantitative RT-PCR and ELISA to study the expression of VEGF mRNA and secretion of VEGF protein. Resuits DADS significantly inhibited proliferation of HL- 60 cell and the inhibiting effects showed a dose （ r 〉 0. 9, P 〈 0.01 ） and time-dependent（ r 〉 0. 7, P 〈 0.01 ） manner. The expression of VEGF mRNA and secretion of VEGF protein could be down regulated by 0.625, 1. 250, and 2. 500 Ixg/mL DADS in HL-60 cells for 24,48 and 72 hours exposure and the effects also showed dose -dependence（r 〉0. 9,P 〈0.01 ）. The growth inhibition rates of DADS in HL-60 cells at three dosages for 24 hours were （8. 19 ± 3.34）%, （ 16.79 ± 2.07）% and （21.30 ± 2.72）%, those for 48 hours were （ 11.93 ± 3.93）%, （22.81 ± 2.31 ）% and （30.74±2.03）%, for 72 hours were （16.68±2.37）%,（28.54±3.26）% and （36.59±2.37）% respectively, The difference between the DADS-treated and untreated HL-60 cells was statistically significant （P 〈 0.01 ）. There were also statistically significant differences among the three groups of different dosages （P 〈 0.01 ）. Conclusion DADS can effectively inhibit the proliferation of HL-60 cells. DADS probably exerts its anti-leukemia effects by reducing the expression of VEGF mRNA and secretion of VEGF protein in HL- 60 cells.