摘要
目的探讨Livin反义寡核苷酸(ASODN)对白血病细胞K562增殖、凋亡和耐药的影响。方法设计合成特异性的Livin硫代磷酸ASODN及其对照错义寡核苷酸(MSODN),脂质体(Lip)介导转染K562细胞,实验分为Lip-ASODN组、Lip-MSODN组、Lip对照组和细胞对照组4组,采用反转录-PCR法检测Livin mRNA的表达;四甲基偶氮唑蓝法与膜联蛋白V异硫氰酸荧光素法检测Livin ASODN与K562细胞增殖、凋亡及与高三尖杉酯碱(HHT)耐药的关系。结果 Livin ASODN可显著抑制K562细胞增殖(P<0.01),且作用具有时间和浓度依赖性。终浓度为600 nmol.L-1的Lip-ASODN能明显降低Livin mRNA表达,细胞凋亡率达(36.89±1.08)%(P<0.01),而Lip-MSODN组、Lip对照组与细胞对照组之间差异均无统计学意义(Pa>0.05)。Livin ASODN可增加K562细胞对HHT的耐药性,半数抑制质量浓度为(0.37±0.02)mg.L-1;与HHT及小剂量阿糖胞苷联合应用可显著抑制细胞增殖(P<0.01)。结论 Livin ASODN可下调Livin基因表达,有效抑制K562细胞的增殖,增加K562细胞凋亡及其对HHT的敏感性。
Objective To investigate effects of Livin antisense oligonucleotide（ASODN） on the proliferation,apoptosis and drug resis-tance of human leukemia（K562）cells.Methods Specific phosphorothioate ASODN and missense oligonucleotide（MSODN） target Livin mRNA were synthesized and transfected into K562 cells following cationic liposome.The experiment was divided into 4 groups： Lip-ASODN group,Lip-MSODN group,Lip control group and cell control group.The expression of Livin mRNA was detected by reverse transcription PCR.The relationship between Livin ASODN and K562 cells was detected by methyl thiazolyl tetrazolium and Annexin V-fluorescein isothiocyanate method.Results The proliferation of K562 cells was inhibited significantly by Livin ASODN in a time-dose-dependent manner（P0.01）.Lip-ASODN at a final concentration of 600 nmol·L-1 could inhibit the expressions of Livin mRNA and the apoptosis rate reached（36.89±1.08）%（P0.01）,but the difference between Lip-MSODN group,Lip control group and cell control group were not statistically significant（Pa0.05）.Livin ASODN could enhance sensitivity to homoharringtonine（HHT） in K562 cells,inhibitory concentration 50 was（0.37±0.02） mg·L-1.Livin ASODN combined with HHT and low-dose Ara-C could inhibit the proliferation of K562 cells apparently（P0.01）.Conclusions Livin ASODN inhibit Livin gene expression and the proliferation of K562 cells effectively,enhances the apoptosis and sensitivity to HHT in K562 cells.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2011年第3期183-186,共4页
Journal of Applied Clinical Pediatrics
