血管内皮生长因子反义寡核苷酸对人白血病细胞血管内皮生长因子表达的影响

VEGF antisense oligonucleotide inhibits the expression of vascular endothelial growth factor in human leukemic cell lines

目的 探讨血管内皮生长因子 (VEGF)反义寡脱氧核苷酸 (AS ODN)对人白血病细胞系VEGF表达的影响。方法 将VEGFAS ODN与K5 6 2和HL 6 0细胞共孵育 ,采用RT PCR技术、免疫组化方法以及ELISA法观察VEGFAS ODN对人白血病细胞系VEGFmRNA和蛋白水平的影响。应用MTT法观察VEGFAS ODN作用后的白血病细胞培养上清对人脐静脉血管内皮细胞 (ECV30 4 )增殖的影响。结果 K5 6 2细胞和HL 6 0细胞经VEGFAS ODN(2 .5～ 15 .0 μmol L)作用 2 4h后 ,VEGFmRNA的表达量呈剂量依赖性减少 ,而其表达量不受VEGF错义ODN的影响 ;当加入 5 .0 μmol LVEGFAS ODN作用 2 4h后 ,细胞内及其培养上清液中VEGF蛋白水平显著减少 ,其培养上清对ECV30 4细胞的促增殖作用减弱 ,而错义ODN处理组白血病细胞VEGF蛋白水平及其作用未见明显改变。结论 VEGFAS ODN在体外能够抑制人白血病细胞VEGF的表达。

Objective To explore the effects of vascular endothelial growth factor ( VEGF ) antisense phosphorothioated oligodeoxynucleotide (AS-ODN) on the expression of VEGF in human leukemic cell lines (HL-60 and K562 cells). Methods The levels of VEGF mRNA and protein in leukemic cells incubated with VEGF AS-ODN were measured by RT-PCR,immunohistochemistry assay and ELISA. MTT test was used to examine the influence of the culture supernatant(CS) of VEGF AS-ODN treated leukemic cells on the proliferation of human umbilical vein endothelial cells (ECV304). Results After leukemic cells were treated with different concentrations (2.5～15.0 μmol/L) of VEGF AS-ODN for 24 h,VEGF mRNA level in the cells decreased remarkably in a concentration dependent manner,no change was found in the VEGF missense ODN treated cells (MS-ODN). When the leukemic cells were treated with 5 μmol/L VEGF AS-ODN for 24 h,VEGF protein level decreased greatly both in the cells and in the CS; and the proliferation stimulating effect of the treated CS on the ECV304 cells reduced. Meanwhile,there was no obvious change in VEGF protein and its effect in the VEGF MS-ODN treated group. Conclusion VEGF AS-ODN could inhibit VEGF expression in human leukemic cell lines in vitro.