Dichloroacetate enhances sorafenib's inhibitory effect on proliferation of hepatocellular carcinoma cells by activating ROS-JNK pathway

Liangbo SUN; Jie YAO; Tao LI; Linxi CHEN; Xiaojing YAN; Fengtian HE; Jiqin LIAN

doi:10.16016/j.1000-5404.201904059

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Dichloroacetate enhances sorafenib's inhibitory effect on proliferation of hepatocellular carcinoma cells by activating ROS-JNK pathway

Basic Medicine | 更新时间：2026-01-09

- Dichloroacetate enhances sorafenib's inhibitory effect on proliferation of hepatocellular carcinoma cells by activating ROS-JNK pathway
- Journal of Third Military Medical University Vol. 41, Issue 17, Pages: 1627-1634(2019)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学):基础医学院生物化学与分子生物学教研室
  2.400038 重庆，陆军军医大学(第三军医大学):生物医学工程与影像医学系数字医学教研室
  3.400037 重庆，陆军军医大学(第三军医大学)第二附属医院全军肿瘤诊治研究所
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81572375);the General Program of Natural Science Foundation of Chongqing(CSTC2018jcyjA2018)
- DOI：10.16016/j.1000-5404.201904059
  CLC： R735.7;R969.2;R979.1
- Received：09 April 2019，
  
  Revised：2019-05-08，
  
  Published：15 September 2019
- 稿件说明：
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Liangbo SUN, Jie YAO, Tao LI, et al. Dichloroacetate enhances sorafenib's inhibitory effect on proliferation of hepatocellular carcinoma cells by activating ROS-JNK pathway[J]. Journal of Third Military Medical University, 2019, 41(17): 1627-1634.
DOI：

Liangbo SUN, Jie YAO, Tao LI, et al. Dichloroacetate enhances sorafenib's inhibitory effect on proliferation of hepatocellular carcinoma cells by activating ROS-JNK pathway[J]. Journal of Third Military Medical University, 2019, 41(17): 1627-1634. DOI： 10.16016/j.1000-5404.201904059.

摘要

目的

探讨二氯乙酸盐(dichloroacetate，DCA)与索拉非尼(sorafenib)联合使用对肝癌细胞Hep3B增殖抑制的效果及可能机制。

方法

将Hep3B细胞分为对照组(DMSO)、DCA处理组(5 mmol/L)、索拉非尼处理组(10 μmol/L)和联合组(5 mmol/L DCA联合10 μmol/L索拉非尼)4个组，处理24 h后，在显微镜下观察各组细胞形态；采用CCK-8检测细胞增殖，流式细胞仪检测细胞凋亡；通过Western blot检测凋亡相关蛋白PARP的表达和p-JNK的水平；采用活性氧(ROS)检测试剂盒检测细胞ROS的变化；用流式细胞仪检测加入抗氧化剂和阻断JNK通路后的细胞凋亡的变化。

结果

DCA和索拉非尼联合处理24 h能够显著改变细胞形态，杀伤细胞。与对照组和单独用药组比较，联合组显著增强对肝癌Hep3B细胞的增殖抑制效果(

0.05)，明显增加Hep3B细胞中PARP的剪切与JNK的磷酸化水平，胞内ROS水平也明显升高；联合使用抗氧化剂NAC可显著抑制DCA和索拉非尼处理导致的JNK磷酸化水平升高和对Hep3B细胞的增殖抑制效果。

结论

DCA和索拉非尼联合使用可显著抑制肝癌细胞Hep3B的增殖，其机制可能与激活ROS-JNK通路有关。

Abstract

Objective

To investigate the synergistic effect of dichloroacetate (DCA) and sorafenib on the inhibition of proliferation in hepatocellular carcinoma (HCC) cells.

Methods

Hep3B cells were treated respectively

with DMSO (control)

DCA (5 mmol/L)

sorafenib (10 μmol/L)

and DCA (5 mmol/L) combined with sorafenib (10 μmol/L) for 24 h

then the cells were stained with crystal violet and observed under a microscope for cell morphology. CCK-8 assay was used to detect the changes of cell proliferation

and flow cytometry was employed to measure cell apoptosis. Then

Western blotting was utilized to detect the protein expression of apoptosis-related protein PARP and p-JNK

and the production of reactive oxygen species (ROS) was measured by ROS detection kits

Finally

flow cytometry was performed to detect the changes of apoptosis after addition of anti-oxidant

NAC and JNK inhibitor

SP600125.

Results

DCA combined with sorafenib significantly changed the cell morphology and killed the cells. Compared with DCA alone or sorafenib alone

DCA combined sorafenib significantly enhanced the inhibitory effect on the proliferation in Hep3B cells (

0.05)

increased the cleavage of PARP and the phosphorylation of JNK. Moreover

the combination induced the production of intracellular ROS (

0.05). Co-treatment of antioxidant NAC resulted in significant inhibition of elevated JNK phosphorylation and inhibitory effect on proliferation induced by DCA combined sorafenib in Hep3B cells.

Conclusion

Combination of DCA and sorafenib significantly inhibits proliferation in Hep3B cells

which may be related with the activation of ROS/JNK signal pathway.

关键词

Keywords

references

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