Survivin反义寡核苷酸对K562细胞耐药性的影响

目的 探讨Survivin反义寡核苷酸(antisense oligodeoxg nucleotide,ASODN)与传统化疗药物对白血病K562细胞耐药性的影响.方法 实验分为反义核苷酸组(ASODN)、无义核苷酸组(SODN)、脂质体组(Lip)、空白对照组,以脂质体转染Survivin ASODN,用RT-PCR检验SurvivinmRNA表达;用MTT法检测K562细胞对阿霉素(driacin,ADM)、柔红霉索(daunorubicin.DAM)及联合用ASODN耐药性;用流式细胞仪检测单独用ADM、DAM和联合用ASODN时K562细胞内ADM和DAM的浓度.结果 ASODN组Survivin mRNA表达较对照组降低(P<0.05);ADM和ASODN ADM对K562细胞的半数抑制细胞(IC50)分别为0.5457 mg/L和0.1933 mg/L,DAM和ASODN DAM对K562细胞的IC5.分别为0.5408 mg/L和0.2027 mg/L,联合用药组ICso低于单用药组(P<0.05);用ADM和ASODN ADM时K562细胞内药物的荧光强度分别为51.64、89.92,DAM和ASODN DAM时K562细胞内药物的荧光强度分别为63.71、88.47,联合用药组细胞内药物浓度高于单用药组(P<0.05).结论 Survivin反义寡核苷酸可下调K562细胞Survivin mRNA的表达;Survivin反义寡核苷酸与阿霉素和柔红霉索联合作用可提高K562细胞内药物浓度,降低细胞生存率,降低细胞耐药性.

Effects of Survivin Antisense Oligodeoxynucleotide on Drug Resistance in K562 Cells

Objective To study the effect of Survivin antisense oligodeoxgnucleotide(ASODN) on drug resistance and its mechanism in K562 cells.Methods Cells were divided into four groups: ASODN group,SODN group,Lip group and blank group.Survivin ASODN was transfered into K562 cells by liposomal reagent.The expression of Survivin mRNA was detected by RT-PCR.The sensitivity of K562 cells to adriamycin(ADM) and daunorubicin(DAM) was detected by MTT assay,while the intracellular concentration of ADM and DAM was measured by flowcytometry.Results Compared to control group Survivin mRNA expression in ASODN group decreased obviously.IC50(s) of ADM and ASODN ADM were 0.5457 mg/L and 0.1933 mg/L respectively,IC50(s) of DAM and ASODN DAM were 0.5408 mg/L and 0.2027 mg/L respectively.Expression of Survivin mRNA decreased by 25.8% after the transfection of ASODN.Fluorescence intensity of ADM and ASODN ADM in K562 cells were 51.64 and 89.92,Fluorescence intensity of DAM and ASODN DAM in K562 cells were 63.71 and 88.47.Conclusion Expression of Survivin mRNA in k562 cells was down-regulated by ASODN.Survivin ASODN is able to reverse the drug resistance via inhibition of Survivin expression and inducement of apoptosis.