天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (12): 1201-1205.doi: 10.11958/20191140

• 细胞与分子生物学 •    下一篇

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

刘迎雪，贾秀红△，尹会颖，朱聪   

  1. 山东滨州，滨州医学院附属医院儿科（邮编256600）
  • 收稿日期:2019-04-12 修回日期:2019-08-26 出版日期:2019-12-15 发布日期:2019-12-15
  • 通讯作者: 贾秀红 E-mail:jiaxiuhong001@163.com
  • 作者简介:刘迎雪（1993），女，硕士在读，主要从事儿童白血病相关研究
  • 基金资助:
    山东省自然科学基金

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

 LIU Ying-xue, JIA Xiu-hong△,YIN Hui-ying, ZHU Cong   

  1. Department of Pediatrics, Binzhou Medical University Hospital, Binzhou 256600, China
  • Received:2019-04-12 Revised:2019-08-26 Published:2019-12-15 Online:2019-12-15
  • Supported by:
     

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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。

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

Abstract: 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.

Key words: drug resistance, neoplasm, leukemia, myeloid, K562 cells, FOXP3, Artesunate, multidug resistance

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