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背景与目的:Survivin是最近发现的凋亡抑制基因,与白血病的发生和耐药有关,反义寡核苷酸可抑制survivin表达,诱导凋亡,增加药物的敏感性。本研究探讨survivin反义寡核苷酸对白血病耐药细胞K562/A02生长、凋亡和半胱氨蛋白水解酶3的活性的作用。方法:以脂质体转染survivin反义寡核苷酸,以MTT、RT-PCR、流式细胞术、荧光分光光度计分别检测survivin反义寡核苷酸对K562/A02细胞的生长抑制、survivin mRNA表达、凋亡和半胱氨蛋白水解酶3的活性。结果:survivin反义寡核苷酸对K562/A02细胞的生长抑制呈浓度-时间依赖性,与对照组相比,survivin mRNA表达降低36.2%(P<0.05),半胱氨蛋白水解酶3的活性增加67.6%(P<0.05);K562/A02细胞的凋亡率明显增加(14.48±2.65%vs 2.39±0.47%,P<0.005)。结论:survivin反义寡核苷酸通过抑制survivin表达,上调半胱氨蛋白水解酶3活性而诱导凋亡。
BACKGROUND & OBJECTIVE: Survivin is a newly discovered inhibitor of apoptosis, which is associated with the development of leukemia and drug resistance. Antisense oligonucleotides inhibit the expression of survivin, induce apoptosis and increase the sensitivity of the drug. This study was to investigate the effect of survivin antisense oligonucleotide on the growth, apoptosis and cysteine protease 3 activity of K562 / A02 cells. Methods: Antisense oligonucleotides of survivin were transfected by liposome. The growth inhibition of K562 / A02 cells was detected by MTT, RT-PCR, flow cytometry and fluorescence spectrophotometer respectively. Survivin mRNA expression, apoptosis and cysteine protease 3 activity. Results: The survivin antisense oligonucleotide inhibited the growth of K562 / A02 cells in a concentration - time dependent manner. Compared with the control group, survivin mRNA expression was decreased by 36.2% (P <0.05), while cysteine protease 3 The activity of K562 / A02 cells increased by 67.6% (P <0.05). The apoptosis rate of K562 / A02 cells increased significantly (14.48 ± 2.65% vs 2.39 ± 0.47%, P <0.005). CONCLUSIONS: Survivin antisense oligonucleotide induces apoptosis by inhibiting survivin expression and up-regulating caspase-3 activity.