SiRNA联合asODN靶向逆转白血病细胞多药耐药的研究  

Reversal of multidrug resistance in leukemia cells by small interfering RNAs combined with antisense oligodeoxyribonucleotides for inhibition of MDR1 gene expression

作　　者：魏军民[1] 侯明[1] 李丽珍[1] 卞继峰[2] 孔峰[2] 

机构地区：[1]山东大学齐鲁医院肿瘤中心,山东济南250012 [2]山东大学医学院生物化学教研室,山东济南250012

出　　处：《山东大学学报（医学版）》2006年第3期283-286,共4页Journal of Shandong University:Health Sciences

摘　　要：目的:探讨靶向多药耐药基因(MDR-1)的siRNA和asODN联合应用逆转人白血病耐药细胞K562/A02的效果。方法:设计并合成针对MDR-1基因同一序列的siRNA和asODN及阴性对照siRNA,采用转染试剂lipofectin2 000分别转染人白血病耐药细胞K562/A02;利用RT-PCR检测MDR-1mRNA和Western Blot检测MDR-1蛋白质的表达;采用罗丹明123外排实验检测P-糖蛋白(P-gp)的转运功能,MTT法检测K562/A02细胞对阿霉素的耐药逆转效果。结果:siRNA、asODNa、sODN和siRNA联合应用均能降低MDR-1mRNA和蛋白质的表达,提高P-gp的转运功能,对阿霉素的敏感性明显恢复,asODN和siRNA联合应用效果明显提高(P<0.05),低浓度的siRNA(200 nmol/L)比高浓度的asODN(5μmol/L)的效果强(P<0.05)。结论:siRNA、asODN能有效地逆转人白血病耐药细胞K562/A02的多药耐药,asODN和siRNA联合应用效果明显加强。Objective： To study the effect of small interfering RNAs（siRNAs）combined with antisense oligodeoxyribonucleotides （asODN） targeted MDR1 Gene on reverting muhidrug resistance of human leukemia cell line K562/A02. Methods： The siRNAs and asODN which targeted the same sequences of MDR1 gene were designed and synthesized. Human leukemia cell line K562/A02 was cultured and tranfected with asODN, siRNAs and negative siRNAs by Lipofectin2000. MDR-lmRNA was assayed by RT-PCR, and the protein expression was detected by Western blotting. The function of P-gp was detected by rhodamine 123 retention, and the efficiency of K562/A02 to ADM was determined by MTT method. Results： The expression of MDR-lmRNA and protein decreased significantly after transfection of asODN, siRNAs ＋ asODN and siRNAs. The transporting function of P-gp increased, and the efficiency of K562/A02 to ADM reversed significantly. The reversing effect increased significantly by siRNAs combined with asODN. The inhibition effect of siRNAs with lower concentration（200 nmol/L）was greater than that of asODN with higher concentration（5 μmol/L）（P 〈 0.05）. Conclusion： Both siRNAs and asODN can reverse multidrug resistance of human leukemia cell line K562/A02. The inhibition effect increases significantly by siRNAs combined with asODN.

关 键 词：小分干扰RNA 寡脱氧核糖核苷酸 反义 基因 MDR 白血病 抗药性 肿瘤 P-糖蛋白 

分 类 号：R733.71[医药卫生—肿瘤]