血管内皮生长因子反义核酸增强HL60和K562细胞对As_2O_3的敏感性

目的:探讨血管内皮生长因子(VEGF)反义核酸能否提高HL60和K562细胞对三氧化二砷(As2O3)的敏感性。方法:采用经筛选所得的最优反义核酸(A7),20个碱基经过全硫代修饰;以脂质体介导转染细胞,反义核酸和As2O3联合作用72h以后,用MTT法检测细胞生长情况,求IC50值;用ELISA法检测培养液中VEGF蛋白的浓度,用流式细胞仪检测细胞凋亡百分数。结果:VEGF反义核酸可显著降低HL60、K562细胞对As2O3的IC50值,下调VEGF蛋白的表达,增加As2O3诱导的HL60、K562细胞凋亡作用。结论:VEGF反义核酸具有增强HL60和K562细胞对As2O3的敏感性,增强As2O3诱导的HL60和K562细胞凋亡作用;提示内源性VEGF蛋白具有使细胞产生耐药性的作用。

VEGF antisense oligodeoxynucleotide increased sensitivity of HL60 and K562 cells to As_2O_3

Aim: To explore the effect of VEGF antisense phosphorothioate oligodeoxynucleotide (AS PS-ODN) on sensitivity of HL60 and K562 cells to As_2O_3. Methods: A7, which was the most effective one of AS PS-ODN selected with help-design of computer and experimental assay, contains 20-mer modified with phosphrothioate and was transferred into cells mediated with lipofectin. 24 hours later, As_2O_3 was added into culture system for continuing incubation for 72 h. And then IC_(50) was determined with MTT method, apoptotic cell numbers with flowcytometry, level of VEGF protein with VEGF ELISA kit. Results: A7 is able to decline IC_(50) level of As_2O_3 of HL60 and K562 cells, to down regulate VEGF protein expression, to increase apoptosis of HL60 and K562 cells induced by As_2O_3. Conclusion:The sensitivity and apoptosis of HL60 and K562 cells to As_2O_3 was enhanced by VEGF AS PS-ODN (A7), which suggested that inner VEGF proteins in leukemia cells bring about side effect of drug-resistance.