沙蟾毒精激活Rho A/ROCK1信号通路诱导急性白血病细胞凋亡的分子机制研究

李志强; 姜秀星; 胡金娇; 丁鑫; 雷令; 高宁

doi:10.16016/j.2097-0927.202109162

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沙蟾毒精激活Rho A/ROCK1信号通路诱导急性白血病细胞凋亡的分子机制研究

药学 | 更新时间：2026-01-09

- 沙蟾毒精激活Rho A/ROCK1信号通路诱导急性白血病细胞凋亡的分子机制研究
- Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway
- 陆军军医大学学报 2022年44卷第7期页码：700-710
- 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：
  
  [高宁，E-mail: gaoning59@163.comGAO Ning, E-mail: gaoning59@163.com
- 基金信息：
  
  国家自然科学基金面上项目(31571425)
- DOI：10.16016/j.2097-0927.202109162
  中图分类号： R733.7;R966;R996.3
- 收稿：2021-09-22，
  
  修回：2021-11-15，
  
  纸质出版：2022-04-15
- 稿件说明：
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李志强, 姜秀星, 胡金娇, 等. 沙蟾毒精激活Rho A/ROCK1信号通路诱导急性白血病细胞凋亡的分子机制研究[J]. 陆军军医大学学报, 2022,44(7):700-710.

Zhiqiang LI, Xiuxing JIANG, Jinjiao HU, et al. Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway[J]. Journal of Army Medical University, 2022, 44(7): 700-710.
李志强, 姜秀星, 胡金娇, 等. 沙蟾毒精激活Rho A/ROCK1信号通路诱导急性白血病细胞凋亡的分子机制研究[J]. 陆军军医大学学报, 2022,44(7):700-710. DOI： 10.16016/j.2097-0927.202109162.

Zhiqiang LI, Xiuxing JIANG, Jinjiao HU, et al. Arenobufagin induces apoptosis in acute leukemia cells by activating Rho A/ROCK1 signaling pathway[J]. Journal of Army Medical University, 2022, 44(7): 700-710. DOI： 10.16016/j.2097-0927.202109162.

摘要

目的

研究沙蟾毒精(arenobufagin

ARE)对急性白血病细胞的凋亡诱导作用及可能的分子机制。

方法

急性白血病细胞Jurkat和MV-4-11经不同浓度(0、10、20、40、80 nmol/L)ARE作用24 h，80 nmol/L ARE作用不同时间(0、6、12、18、24 h)或ROCK1抑制剂Y-27632(20 μmon/L)预处理1 h后80 nmol/L ARE作用24 h，采用MTT检测ARE抑制急性白血病细胞增殖的能力；采用流式细胞术检测ARE诱导急性白血病细胞凋亡的能力；采用Western blot和CoIP实验检测急性白血病细胞经ARE作用后细胞中凋亡相关蛋白和Rho A/ROCK1信号通路相关蛋白的变化情况。

结果

ARE对Jurkat和MV-4-11细胞的抑制增殖作用呈剂量依赖性和时间依赖性(

＜0.01)，对Jurkat和MV-4-11细胞凋亡诱导作用亦呈剂量和时间依赖性(

＜0.01)；Western blot检测结果显示: ARE可激活ROCK1信号通路，促进BAX移位至线粒体，Cytochrome c (Cyto c)和AIF由线粒体释放入细胞质，增加Caspase 3和Caspase 7剪切激活，增加PARP1剪切，并减少XIAP表达，其上述作用具有剂量依赖性和时间依赖性(

＜0.01)；使用Y-27632抑制ROCK1信号通路可明显阻断ARE诱导的BAX移位至线粒体、Cyto c和AIF向细胞质的释放、Casp

ase 3和Caspase 7的剪切激活、PARP1的剪切、XIAP表达的下调以及细胞凋亡(

＜0.01)。

结论

ARE通过激活Rho A/ROCK1信号通路有效诱导急性白血病细胞(Jurkat、MV-4-11)发生凋亡。

Abstract

Objective

To investigate the apoptosis-inducing effect of arenobufagin (ARE) on acute leukemia cells and explore its possible molecular mechanism.

Methods

Acute leukemia cell lines Jurkat and MV-4-11 were treated with different concentrations of ARE (0

40 or 80 nmol/L) for 24 h

or with 80 nmol/L ARE for different durations (0

18 or 24 h)

or with 80 nmol/L ARE for 24 h after being pretreated with 20 μmol/L Y-27632 (inhibitor of ROCK1) for 1 h

respectively. Then

MTT assay and flow cytometry were used to respectively detect the effect of ARE on cell proliferation and apoptosis in acute leukemia cells; Western blotting and co-immunoprecipitation (CoIP) assay were performed to determine the expression of apoptosis-related and Rho A/ROCK1 pathway-related proteins in the acute leukemia cells after ARE treatment.

Results

ARE inhibited the proliferation and induced apoptosis of acute leukemia cells in dose- and time-dependent manners (

0.01). Western blotting results showed that ARE activated ROCK1 signaling pathway

facilitated the translocation of BAX from cytoplasm to mitochondria

promoted the release of mitochondrial proteins (Cytochrome c

AIF)

increased the cleavage/activation of Caspase 3 and Caspase 7 as well as the degradation of Poly(ADP-ribose) polymerase 1 (PARP1)

and then reduced the expression of X-linked inhibitor of apoptosis (XIAP). The above effects presented dose- and time-dependence (

0.01). Interruption of ROCK1 pathway by the pharmacological inhibitor Y-27632 significantly attenuated ARE-mediated mitochondrial translocation of BAX

reduced cytosolic release of Cytochrome c and AIF from mitochondria

ppressed the activation of Caspase 3 and Caspase 7

the degradation of cleavage of PARP1

and down-regulated XIAP expression as well as blocked apoptosis (

0.01).

Conclusion

ARE effectively induces apoptosis of acute leukemia cells (Jurkat

MV-4-11) through the activation of Rho A/ROCK1 signaling pathway.

关键词

Keywords

references

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