Survivin反义寡核苷酸对5-FU诱导人红白血病细胞系K562凋亡的影响

Survivin antisense oligodeoxy-nucleotid enhances 5-FUinduced apoptosis of leukemic cell line K562

【目的】探讨survivin反义寡核苷酸(Antisense Oligodeoxy-nucleotid,ASODN)对5-FU(5-氟尿嘧啶)诱导人红白血病细胞系K562细胞凋亡的影响。【方法】体外培养K562细胞,合成survivin ASODN并经脂质体转染至K562细胞内,MTT法观察survivin ASODN组、5-FU组及5-FU联合survivin ASODN的细胞毒作用,Hoechst33342/PI双荧光染色观察各组细胞核形态,镜下计算各组细胞凋亡率。【结果】400、600、800、1000nmol/L survivin ASODN处理K562细胞44 h后,IC50为800 nmol/L;与单独使用survivin ASODN或5-FU相比,5-FU联合800 nmol/L survivin ASODN后细胞生长明显受到抑制(P<0.01);Hoechest33342/PI双荧光染色可观察到survivin ASODN组、5-FU组及5-FU联合survivin ASODN组均出现明显核固缩、凝集等细胞凋亡表现。镜下细胞计数,survivin ASODN组、5-FU组及5-FU联合survivin ASODN组细胞凋亡率分别为54.55%、53.85%、86.70%。【结论】Survivin ASODN可增强K562细胞对5-FU的敏感性。

[Objective] To investigate the effects of survivin antisense ollgodeoxy-nucleotid （ASODN） on 5-FU- induced apoptosis in leukemic cell llne K562. [Methods] Survivin ASODN was transferred into K562 by lipofectin, and the cytotoxicity of survivin ASODN were examined by MTT assay. Morphological changes of the nucleous were observed by Hoechst33342/PI staining. [Results] The IC50 for K562 cells was 800 nmoL/L. When K562 cells were treated with survivin ASODN at the concentration of 400, 600, 800 and 1 000 nmoL/L the growth of cell was inhibited remarkably with dosage dependency. The inhibition effects of 5-FU ＋ ASODN group were remarkable as compared with the simple ASODN or simple 5-FU group.. Distinct morphological changes of cell apoptosis such as karyopyknosls and conglomeration were observed by Hoechst33342/PI staining. The apoptosis rate of survivin ASODN group, 5-FU group and 5-FU combinated with survivin ASODN group was 54.55%, 53.85% and 86.70% repectively. [Conclusion] Survivin ASODN could enhance 5-FU -induced apoptosis in leukemic cell line K562.