Survivin反义寡核苷酸诱导K562细胞株凋亡的实验研究

Apoptosis of K562 cells induced by antisense oligonucleotide targeting Survivin

目的探讨Survivin反义寡核苷酸对K562细胞株凋亡的影响。方法将K562细胞分为四组:反义组、无义组、脂质体组及空白对照组,通过脂质体将Survivin反义寡核苷酸转染入各组K562细胞,用免疫组化法测定转染前后K562细胞Survivin基因及Caspase-3的表达差异,用Tunel及Annexin V-FITC测定各组细胞的凋亡率。结果Survivin基因在K562细胞高表达,转染反义核苷酸后其表达下降,转染Survivin基因反义核苷酸后Caspase-3的表达较其它各组升高,K562细胞生长受抑,细胞凋亡率升高。结论Survivin反义寡核苷酸能够下调Survivin基因的表达,促进白血病细胞株K562的凋亡。

Objective To study the apoptosis of leukemia cells induced by antisense oligonucleotide targeting Survivin. Methods Oligonucleotides were delivered in the form of complexes with Lipofectin. K562 cells were divided into four groups： ASON group, NSON group, lipofectin group, and control group. The expression of Survivin Caspse-3 was examined by immunohistochemistry（ SABC ）. And detected the apoptotic rate of each group with two methods ： Tunel assay and Annexin V-FITC assay. Results The protein of Survivin is overexpress in K562 cells ,but after treated with antisense oligonucleotide, the level of expression was down regulation. After transfection, the expression of caspase-3 of ASON group was upgrade contrast to other three groups （ P 〈 0.05 ）. The apoptotic rates detected by Tunel assay and Annexin V-FITC assay of ASON group were all higher than other three groups （ P 〈 0.05 ）. Conclusion Antisense oligonucleotide targeting Survivln can down regulate the protein expression of Survivin, and can induce K562 cells to apoptosis.