青蒿琥酯抑制FOXP3基因对K562/ADR细胞增殖、凋亡及多药耐药的影响  

Effects of artesunate inhibiting the expression of FOXP3 on proliferation, apoptosis and multidrug resistance of K562/ADR cells

作　　者：刘迎雪 贾秀红[1] 尹会颖 朱聪[1] LIU Ying-xue;JIA Xiu-hong;YIN Hui-ying;ZHU Cong(Department of Pediatrics,Binzhou Medical University Hospital,Binzhou 256600,China)

机构地区：[1]山东滨州滨州医学院附属医院儿科

出　　处：《天津医药》2019年第12期1201-1205,共5页Tianjin Medical Journal

摘　　要：目的探讨青蒿琥酯(Artesunate)抑制FOXP3基因表达对慢性髓系白血病(CML)耐阿霉素(ADR)细胞株K562/ADR增殖、凋亡及多药耐药的影响,并探讨其作用机制。方法实时荧光定量PCR(Real-time PCR)技术检测FOXP3 mRNA在K562和K562/ADR细胞中的表达;蛋白印迹法(Western blot)检测FOXP3蛋白的表达;青蒿琥酯不同质量浓度(2.5、5.0、7.5、10.0、12.5 mg/L)处理K562/ADR细胞24 h,利用CCK-8法检测不同浓度青蒿琥酯对K562/ADR细胞的细胞毒性,筛选无细胞毒性浓度的青蒿琥酯完成后续实验;RT-PCR法、Western blot法检测无细胞毒性浓度青蒿琥酯处理下FOXP3 mRNA、FOXP3蛋白表达变化;CCK-8法检测ADR对细胞毒性的变化;流式细胞术(FCM)检测ADR平均荧光强度,即蓄积浓度变化。结果 FOXP3基因在CML耐药细胞株K562/ADR中表达升高;无毒剂量浓度青蒿琥酯(2.5、5.0、7.5 mg/L)处理下K562/ADR细胞FOXP3 mRNA、蛋白表达受抑制(P<0.05);K562/ADR细胞胞内ADR毒性增强,浓度升高(P<0.05)。结论 FOXP3基因在CML K562/ADR耐药细胞株中高表达;青蒿琥酯可以通过抑制FOXP3基因表达,增加K562/ADR胞内ADR浓度,逆转多药耐药,且呈一定的剂量依赖性。Objective To investigate the effect of artesunate inhibiting the expression of FOXP3 on proliferation,apoptosis and multidrug resistance of adriamycin(ADR)-resistant K562/ADR cells in chronic myeloid leukemia(CML),and to explore its mechanism.Methods The expressions of FOXP3 mRNA in K562 and K562/ADR cells were detected by real-time PCR.The expressions of FOXP3 proteins in K562 and K562/ADR cells were detected by Western blot assay.The K562/ADR cells were treated with different concentrations of artesunate(2.5,5.0,7.5,10.0 and 12.5 mg/L) for 24 h.The toxicities of different concentrations of artesunate to K562/ADR cells were detected by CCK-8 assay,and the non-cytotoxic concentrations were screened.The expressions of FOXP3 mRNA and proteins in K562/ADR cells treated by non-cytotoxic concentration of artesunate were detected by RT-PCR and Western blot assay.The changes of toxicities of ADR in K562/ADR cells were detected by CCK-8 assay.The average fluorescence intensities of ADR were detected by FCM assay.Results The expressions of FOXP3 were higher in K562/ADR cells than those in K562 cells.The mRNA and proteins expressions of FOXP3 were significantly lower in 2.5 mg/L group,5 mg/L group and 7.5 mg/L group than those in the control group.The toxicities and concentrations of ADR were increased in K562/ADR cells treated by artesunate(both P<0.05).Conclusion FOXP3 gene is highly expressed in adriamycin-resistant K562/ADR cells in CML.Artesunate can increase the concentrations of ADR and reverse multidrug resistance in K562/ADR cells by inhibiting the expression of FOXP3 in a dose-dependent manner.

关 键 词：抗药性 肿瘤 白血病 髓样 K562细胞 FOXP3基因 青蒿琥酯 多药耐药 

分 类 号：R733.7[医药卫生—肿瘤]