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

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。

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

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, semi-quantitative RT-PCR and ELISA to study the expression of VEGF mRNA and secretion of VEGF protein. RESULTS: 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 microg/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.