Molecular mechanism of oprozomib synergistic with arsenic trioxide in inhibiting proliferation of acute promyeloid leukemia cells

Xin DING; Xiuxing JIANG; Zhiqiang LI; Ling LEI; Ning GAO

doi:10.16016/j.2097-0927.202109071

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Molecular mechanism of oprozomib synergistic with arsenic trioxide in inhibiting proliferation of acute promyeloid leukemia cells

Basic Medicine | 更新时间：2026-01-09

- Molecular mechanism of oprozomib synergistic with arsenic trioxide in inhibiting proliferation of acute promyeloid leukemia cells
- Journal of Army Medical University Vol. 44, Issue 4, Pages: 329-336, 355(2022)
- 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与中药学教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(31571425)
- DOI：10.16016/j.2097-0927.202109071
  CLC： R733.71;R966;R979.19
- Received：08 September 2021，
  
  Revised：2021-11-07，
  
  Published：28 February 2022
- 稿件说明：
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Xin DING, Xiuxing JIANG, Zhiqiang LI, et al. Molecular mechanism of oprozomib synergistic with arsenic trioxide in inhibiting proliferation of acute promyeloid leukemia cells[J]. Journal of Army Medical University, 2022, 44(4): 329-336, 355.
DOI：

Xin DING, Xiuxing JIANG, Zhiqiang LI, et al. Molecular mechanism of oprozomib synergistic with arsenic trioxide in inhibiting proliferation of acute promyeloid leukemia cells[J]. Journal of Army Medical University, 2022, 44(4): 329-336, 355. DOI： 10.16016/j.2097-0927.202109071.

摘要

目的

探讨奥普佐米协同三氧化二砷抑制急性早幼粒白血病细胞增殖分子机制。

方法

MTT法检测不同联合用药对急性早幼粒白血病NB4、HL-60细胞存活率的影响: ①梯度浓度三氧化二砷(NB4细胞: 0、0.1、0.2、0.3、0.4、0.5 μmol/L；HL-60细胞: 0、0.4、0.8、1.2、1.6、2.0 μmol/L)单用或与固定浓度奥普佐米(NB4细胞: 40 nmol/L；HL-60细胞: 160 nmol/L)联用；②梯度浓度奥普佐米(NB4细胞: 0、5、10、20、40 nmol/L；HL-60细胞: 0、40、80、120、160、200 nmol/L)单用或与固定浓度三氧化二砷(NB4细胞: 0.4 μmol/L；HL-60细胞: 1.2 μmol/L)联用；③单独或联合用药作用不同时间(12、24、48 h)；④相同浓度的硼替佐米或奥普佐米分别与梯度浓度三氧化二砷联用。单独用药或联合用药或用NAC预处理后，流式细胞仪检测细胞凋亡及ROS生成量；Western blot检测凋亡相关蛋白cleaved-Caspase 3、cleaved-Caspase 9、PARP-1及融合基因蛋白PML-RARA的表达。

结果

与单用组比较，奥普佐米与三氧化二砷联用后，NB4、HL-60细胞存活率显著降低(

＜0.01)，呈剂量和时间依赖性，并具有明显的协同作用，协同系数小于1。联合用药可使约80%的NB4细胞ROS水平明显升高，融合蛋白PML-RARA表达降低，细胞凋亡率增加约70%(

＜0.01)；与联合用药组比较，ROS抑制剂NAC预处理后，细胞凋亡减少约41%(

＜0.01)，PML-RARA的表达升高。

结论

奥普佐米协同三氧化二砷诱导急性早幼粒白血病细胞ROS生成，并导致PML-RARA蛋白表达降低，促进细胞凋亡。

Abstract

Objective

To investigate the molecular mechanism of oprozomib synergistically acting with arsenic trioxide in inhibiting the proliferation of acute promyeloid leukemia cells.

Methods

MTT assay was used to observe the effects of different medication strategies on the survival rate of NB4 and HL-60 cells respectively: ①gradient concentration of arsenic trioxide was adopted alone (for NB4 cells: 0

0.1

0.2

0.3

0.4

0.5 μmol/L; for Hl-60 cells: 0

0.4

0.8

1.2

1.6

2.0 μmol/L) or combined with fixed concentration of oprozomib (for NB4 cells: 40 nmol/L; for Hl-60 cells: 160 nmol/L); ②gradient concentration of oprozomib was applied alone (for NB4 cells: 0

40 nmol/L; for Hl-60 cells: 0

120

160

200 nmol/L) or combined with fixed concentration of arsenic trioxide (for NB4 cells: 0.4 μmol/L; for Hl-60 cells: 1.2 μmol/L); ③ the treatment results of single drug or combination were observed for 12

24 and 48 h respectively; ④the effects of bortezomib or oprozomib (both at the same concentration) were compared when in combination with arsenic trioxide (at gradient concentration). In addition

flow cytometry was used to detect cell apoptosis and reactive oxygen species (ROS) production

and Western blotting was performed to determine the expression of apoptosis related proteins cleaved-Caspase 3

cleaved-Caspase 9

PARP-1 and the fusion gene protein PML-RARA

in the presence or absence of N-acetylcysteine (NAC

ROS inhibitor

5 mmol/L).

Results

As compared with the single drug group

the combination of oprozomib and arsenic trioxide resulted in a significant decline of cell viability of NB4 and HL-60 cells (

0.01) in a dose- and time-dependent manner

showing an obvious synergistic effect

and the synergistic coefficient was less than 1. The combined medication strategy also increased the ROS production by about 80%

improved the apoptotic rate by 70%

and reduced the expression of PML-RARA in NB4 cells(

0.01). However

inhibiting ROS production by NAC pretreatment decreased the apoptotic rate by about 41% (

0.01)

along with elevated expression of PML-RARA.

Conclusion

The combination of oprozomib and arsenic trioxide promotes apoptosis by inducing ROS generation and reducing the expression of PML-RARA protein in acute promyelocytic leukemia cells.

关键词

Keywords

references

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