Molecular mechanism of cepharanthine synergistically acting with doxorubicin in reversing multidrug resistance of K562/ADR cells

Xiuxing JIANG; Ruoqiu FU; Ning GAO

doi:10.16016/j.1000-5404.201807166

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Molecular mechanism of cepharanthine synergistically acting with doxorubicin in reversing multidrug resistance of K562/ADR cells

Basic Medicine | 更新时间：2026-01-09

- Molecular mechanism of cepharanthine synergistically acting with doxorubicin in reversing multidrug resistance of K562/ADR cells
- Journal of Third Military Medical University Vol. 40, Issue 23, Pages: 2154-2161(2018)
- 作者机构：
  
  400038 重庆, 陆军军医大学(第三军医大学)药学与检验医学系生药学教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(31571425)
- DOI：10.16016/j.1000-5404.201807166
  CLC： R285.5;R733.7;R915
- Received：23 July 2018，
  
  Revised：2018-08-24，
  
  Published：15 December 2018
- 稿件说明：
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Xiuxing JIANG, Ruoqiu FU, Ning GAO. Molecular mechanism of cepharanthine synergistically acting with doxorubicin in reversing multidrug resistance of K562/ADR cells[J]. Journal of Third Military Medical University, 2018, 40(23): 2154-2161.
DOI：

Xiuxing JIANG, Ruoqiu FU, Ning GAO. Molecular mechanism of cepharanthine synergistically acting with doxorubicin in reversing multidrug resistance of K562/ADR cells[J]. Journal of Third Military Medical University, 2018, 40(23): 2154-2161. DOI： 10.16016/j.1000-5404.201807166.

摘要

目的

探讨千金藤素协同多柔比星逆转K562/ADR细胞耐药的分子机制。

方法

MTT法测定K562、K562/ADR细胞存活率; Western blot检测多药耐药蛋白Mdr-1、凋亡相关蛋白以及线粒体分裂关键蛋白Drp1和p-Drp1(Ser637)、线粒体外膜蛋白Tom20蛋白的表达:流式细胞仪检测K562/ADR细胞凋亡以及ROS生成量; 免疫共沉淀技术检测Drp1、Tom20蛋白之间特异性结合。

结果

梯度浓度多柔比星均会降低K562、K562/ADR细胞存活率

两者的IC

值分别为(0.75±0.11)、(75.97±2.17)μmol/L

差异有统计学意义(

0.01)。K562细胞中检测不到多药耐药蛋白

而在K562/ADR细胞中Mdr-1高表达。千金藤素与多柔比星协同作用于K562/ADR细胞

导致K562/ADR存活率明显降低并呈较好的量效关系

协同系数均小于1

K562/ADR细胞凋亡率显著增加(

0.01)。千金藤素与多柔比星协同作用于K562/ADR细胞导致ROS生成量显著升高

Drp1线粒体转位增加。抑制ROS则可阻断两种药物协同作用引起的Drp1线粒体转位

降低K562/ADR细胞凋亡率(

0.01)。

结论

千金藤素协同多柔比星诱导ROS生成促进Drp1线粒体转位

导致线粒体过度分裂和细胞凋亡

最终逆转K562/ADR耐药。

Abstract

Objective

To investigate the molecular mechanism of cepharanthine synergistically acting with doxorubicin in the reverse of multidrug resistance of K562/ADR cells.

Methods

After K562 cells and K562/ADR cells were treated with doxorubicin at various concentration respectively

cell viability was assessed by MTT assay. Then cepharanthine (6 μmol/L)

doxorubicin (4 μmol/L)

NAC (ROC inhibitor

4 μmol/L) alone

or in combination of the former 2 agents or 3 agents together were used to treat the cells respectively. Cell apoptosis and production of reactive oxygen species (ROS) were determined by flow cytometry with Annexin V-/PI staining. The expression of multidrug resistant protein Mdr-1

apoptosis and mitochondrial fission related proteins

Drp1 and p-Drp1 (Ser637)

and translocase of the mitochondrial outer membrane Tom20 were detected by Western blot analysis. Co-immunoprecipitation was used detected the binding of Drp1 and Tom20.

Results

The IC

value of K562 cells to doxorubicin was 0.75±0.11 μmol/L

and that of K562/ADR cells was 75.97±2.17 μmol/L

with significantly difference (

0.01). Mdr-1 was mildly expressed in K562 cells

but was highly expressed in K562/ADR cells. Combined treatment of cepharanthine and doxorubicin resulted in marked decrease in cell viability

which was in a dose-dependent manner and the synergistic coefficient less than 1

and also resulted in increases in apoptotic rate (

0.01) in K562/ADR cells. The synergistic action of the 2 agents also promoted the production of ROS and mitochondrial translocation of Drp1. What's more

inhibiting ROS production by NAC treatment could block the mitochondrial translocation of Drp1 and decrease the apoptotic rate induced by the 2 agents (

0.01).

Conclusion

Cepharanthine synergistically acting with doxorubicin induces the production of ROS and promotes mitochondrial translocation of Drp1

thus results in mitochondrial fission and cell apoptosis

and finally reverses multidrug resistance of K562/ADR cells.

关键词

Keywords

references

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