siRNA沉默c-FLIP对K562/ADR耐药性的影响  

Effect of si RNA-mediated silencing of c-FLIP on adriamycin-resistance of K562/ADR cells

作　　者：宋敏[1] 王建宁[1] 孟庆齐[1] 包红雨[1] 杨杰[2] 

机构地区：[1]南京医科大学第二附属医院血液科,江苏南京210011 [2]江苏省中医药研究院,江苏南京210028

出　　处：《海南医学》2015年第11期1561-1565,共5页Hainan Medical Journal

基　　金：南京医科大学科技发展基金重点项目(编号:2010NJMUZ49)

摘　　要：目的探讨c-FLIP si RNA干扰c-FLIP m RNA水平对阿霉素耐药细胞K562/ADR的耐药性的影响及作用机制。方法应用si RNA干扰的方法抑制K562及K562/ADR细胞c-FLIP的表达,通过荧光定量PCR的方法检测c-FLIP m RNA表达及对多药耐药基因MDR1 m RNA水平的影响。MTT法检测c-FLIP干扰与否对K562及K562/ADR细胞增殖的影响,Annexin V/7-ADD双染研究c-FLIP干扰与否对K562及K562/ADR细胞凋亡的影响。结果与阴性si RNA转染组相比较,c-FLIP si RNA转染下调K562细胞中c-FLIP的m RNA后,K562细胞增殖受到一定程度的抑制(P<0.05),但是并未显著诱导细胞凋亡,c-FLIP干扰与否K562细胞48 h增殖率分别为(69.14±1.82)%和(60.69±2.23)%,凋亡率分别为(1.7±0.3)%和(1.8±0.2)%。与阴性si RNA转染组相比较,c-FLIP si RNA转染抑制K562/ADR细胞中c-FLIP的m RNA后,K562/ADR细胞增殖显著被抑制(P<0.05),并显著诱导了细胞凋亡增加(P<0.05),c-FLIP干扰与否K562/ADR细胞48 h增殖率分别为(-6.07±0.71)%和(-37.45±3.53)%,凋亡率分别为(5.2±0.4)%和(9.2±0.4)%。并且c-FLIP si RNA下调c-FLIP m RNA水平后,K562/ADR细胞中的多药耐药基因MDR1的m RNA表达水平也被显著下调(P<0.05)。结论 c-FLIP si RNA下调K562/ADR细胞中c-FLIP的m RNA水平抑制了多药耐药基因MDR1的表达,从而抑制了K562/ADR细胞对阿霉素的耐药性。Objective To investigate the effect of silenced c-FLIP mRNA level by small interfering RNA （siRNA） on adriamycin-resistance of K562/ADR cells. Methods c-FLIP siRNA and negative siRNA were transfect-ed into K562 and K562/ADR cell lines respectively, and mRNA expression of c-FLIP and multi-drug resistance gene 1 （MDR1） were detected by quantitative PCR. Cell proliferation rate was detected by MTT assay, and cell apoptosis rate was assayed by Annexin V/7-ADD double-staining method. Results Compared with negative siRNA transfection group, siRNA transfection significantly decreased c-FLIP mRNA expression in K562 cells （P〈0.05） and slightly inhib-ited the proliferation of K562 cell （not enough to induce K562 cell apoptosis）. The 48 h proliferation rates of K562 cells were （69.14 ± 1.82）% and （60.69 ± 2.23）% in negative siRNA transfection group and siRNA transfection group, and the apoptosis rate were （1.7±0.3）%and （1.8±0.2）%, respectively. c-FLIP siRNA transfection significantly inhibit-ed K562/ADR proliferation and induced cell apoptosis （P〈0.05）. The 48 h proliferation rate of K562/ADR cells were （-6.07 ± 0.71）% and （-37.45 ± 3.53）% in negative siRNA transfection group and siRNA transfection group, and the apoptosis rate were （5.2 ± 0.4）% and （9.2 ± 0.4）%, respectively. In addition, MDR1 mRNA expression of K562/ADR was significantly decreased （P〈0.05） upon c-FLIP siRNA transfection （P〈0.05）. Conclusion c-FLIP siRNA transfec-tion significantly decreases mRNA expression of MDR1 and inhibites the adriamycin-resistance of K562/ADR cells.

关 键 词：C-FLIP SIRNA MDR1 K562/ADR 阿霉素耐药 

分 类 号：R96[医药卫生—药理学]