重组人血管内皮抑素对体外培养血管瘤内皮细胞增殖、细胞周期及VEGF、KDR／FIK-1表达的影响

摘要

Objective To observe inhibitory effect of recombinant human endostatin on the proliferation of vascular tumor endothelial cells and to investigate its possible mechanism.Methods Hemangioma endothelial cells were cultured in vitro and different concentrations of endostatin on hemangioma endothelial cell proliferation inhibition were detected by MTT method.Effect of recombinant human endostatin on endothelial cell cycle was detected by flow cytometry.The expression of VEGF, KDR mRNA in hemangioma endothelial cell were detected by Real-time RT PCR. Results Recombinant human endostatin concentrations in 24 h, 48 h and 72 h after three period during which the hemangioma endothelial cells inhibited significantly( P <0.01 ) , and there was a clear dose dependence, IC50 was 355 μg/mL.Recombinant human vascular endostatin ( 250 μg/mL ) intervented for 24 hours, the proportion of cells in G0/G1 phase(94.23 ±1.66)%, compared with control group (90.63 ±1.14)%, had significantly difference (P<0.05).Compared with the control group, the difference of the expression of vascular endothelial growth factor (VEGF) was statistically significant (P<0.05) as well as Flk-1 (P <0.05).Conclusion Recombinant human endostatin does not only have inhibitory effect of hemangioma endothelial cell cycle, but also can inhibit the expression of VEGF and FLK-1.%目的：观察重组人血管内皮抑制素对血管瘤内皮细胞增殖的抑制作用，探讨其可能的作用机制。方法体外培养的血管瘤内皮细胞受到重组人血管内皮抑制素的作用后，采用MTT法检测血管瘤内皮细胞增殖，流式细胞术检测血管瘤内皮细胞周期，实时荧光定量PCR技术检测VEGF、KDR mRNA在血管瘤内皮细胞中的表达。结果重组人血管内皮抑制素各浓度组在24、48、72 h 3个作用时段对血管瘤内皮细胞抑制作用显著（P＜0.01），并存在明显的剂量依赖关系，IC50＝355μg／mL；重组人血管内皮抑制素（250μg／mL）干预24 h后，G0／G1期细胞比例（94.23±1.66）％较对照组（90.63±1.14）％有明显增多，差异有统计学意义（P＜0.05）；VEGF与Flk-1 mRNA的表达相比对照组明显降低（ P＜0.05）。结论血管瘤内皮细胞周期受到重组人血管内皮抑制素抑制作用，同时VEGF、FLK-1的表达也受到其有效抑制。