生存素反义寡核苷酸诱导HL-60细胞凋亡的作用探讨

Effect of antisense oligodeoxynucleotide(ASODN)targeting survivin on inducing apoptosis of HL-60 cells

目的:探讨特异性生存素(survivin)反义寡核苷酸(ASODN)对急性早幼粒细胞系HL-60细胞增殖、凋亡的影响。方法:人工合成survivin硫代ASODN,通过脂质体转染HL-60细胞后,采用MTT法检测细胞增殖抑制作用,倒置显微镜观察细胞形态学变化,原位末端标记(Tunel)法检测细胞凋亡指数(AI),RT-PCR半定量测定细胞中survivinmRNA的表达。结果:(1)survivinASODN各浓度组对HL-60细胞的增殖均有抑制作用,且呈时间-剂量依赖性。(2)survivinASODN处理组HL-60细胞的凋亡率与空白对照组、正义对照组和空转染组相比,差异有显著性(P<0.01)。(3)survivinASODN处理组HL-60细胞的survivinmRNA表达水平明显下调。结论:survivinASODN能明显抑制HL-60细胞中survivinmRNA的表达,诱导细胞凋亡。

Objective To explore the effects of ASODN targeting survivin on the proliferation and apoptosis of HL-60 cells. Methods Phosphorothioated ASODN targeting survivin was synthesized and transfected into HL-60 cells by lipofectin, MTT assay was used to detect the inhibitory effect on the cell proliferation. Morphological changes of the cells were observed under an inverted microscope. Apoptotic index was examined by Tune1. The expression of survivin mRNA was detected by reverse transcriptase PCR. Results （1） 200 - 1 000 nmol/L ASODN targeting surviving had an obvious inhibitory effect on the proliferation of HL-60 cells and this effect was correlated with time and dosage. （2）The apoptotic rate in ASODN targeting survivin group was markedly higher than that in three control groups （ P〈0.01 ）. （3）The expression of survivin mRNA significantly decreased after treatment with ASODN targeting survivin. Conclusion ASODN targeting survivin can effectively inhibit the expression of survivin mRNA and induce the apoptosis of HL-60 cells.