标题：siRNA沉默c-FLIP对K562/ADR耐药性的影响

      作者：宋 敏 1，王建宁 1，孟庆齐 1，包红雨 1，杨 杰 2
    (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)

      卷次： 2015年26卷11期

      【摘要】 目的 探讨 c-FLIP siRNA干扰 c-FLIP mRNA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 siRNA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 mRNA表达及对多药耐药基因MDR1 mRNA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 siRNA转染组相比较，c-FLIP siRNA转染下调K562细胞中 c-FLIP的mRNA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 siRNA转染组相比较，
c-FLIP siRNA转染抑制K562/ADR细胞中c-FLIP的mRNA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 siRNA下调c-FLIP mRNA水平后，K562/ADR细胞中的多药
耐药基因MDR1的mRNA表达水平也被显著下调(P<0.05)。结论 c-FLIP siRNA下调K562/ADR细胞中 c-FLIP
的mRNA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。

      【关键词】 c-FLIP siRNA；MDR1；K562/ADR；阿霉素耐药

      【中图分类号】 R96 【文献标识码】 A 【文章编号】 1003—6350（2015）11—1561—05


Effect of siRNA-mediated silencing of c-FLIP on adriamycin-resistance of K562/ADR cells.
SONG Min 1, WANG
Jian-ning 1, MENG Qing-qi 1, BAO Hong-yu 1, YANG Jie 2. 1. Department of Hematology, the Second Affiliated Hospital
of Nanjing Medical University, Nanjing 210011, Jiangsu, CHINA; 2. Jiangsu Province Institute of Traditional Chinese
Medicine, Nanjing 210028, Jiangsu, CHINA

【Abstract】 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.

      【Key words】 Small interfering RNA (siRNA); Multi-drug resistance gene 1 (MDR1); K562/ADR; Adriamycin
resistance
基金项目：南京医科大学科技发展基金重点项目(编号：2010NJMUZ49)