[1]孙梁博,姚洁,李涛,等.二氯乙酸盐激活ROS-JNK通路增强索拉非尼对肝癌细胞增殖的抑制作用[J].第三军医大学学报,2019,41(17):1627-1634.
 SUN Liangbo,YAO Jie,LI Tao,et al.Dichloroacetate enhances sorafenib’s inhibitory effect on proliferation of hepatocellular carcinoma cells by activating ROS-JNK pathway[J].J Third Mil Med Univ,2019,41(17):1627-1634.
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二氯乙酸盐激活ROS-JNK通路增强索拉非尼对肝癌细胞增殖的抑制作用(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第17期
页码:
1627-1634
栏目:
基础医学
出版日期:
2019-09-15

文章信息/Info

Title:
Dichloroacetate enhances sorafenib’s inhibitory effect on proliferation of hepatocellular carcinoma cells by activating ROS-JNK pathway
作者:
孙梁博姚洁李涛陈岺曦闫小晶何凤田连继勤
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1,生物医学工程与影像医学系数字医学教研室2;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3
Author(s):
SUN Liangbo YAO Jie LI Tao CHEN LinxiYAN Xiaojing HE Fengtian LIAN Jiqin

Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, 2Department of Digital Medicine, Faculty of Biomedical Engineering and Imaging Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038; 3Cancer Institute, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China

关键词:
二氯乙酸盐索拉非尼肝癌细胞增殖ROS-JNK通路
Keywords:
dichloroacetate sorafenib hepatocellular carcinoma cell proliferation ROS/JNK signal pathway
分类号:
R735.7; R969.2; R979.1
文献标志码:
A
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 (P<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:

ObjectiveTo investigate the synergistic effect of dichloroacetate (DCA) and sorafenib on the inhibition of proliferation in hepatocellular carcinoma (HCC) cells. MethodsHep3B 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. ResultsDCA 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 (P<0.05), increased the cleavage of PARP and the phosphorylation of JNK. Moreover, the combination induced the production of intracellular ROS (P<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. ConclusionCombination of DCA and sorafenib significantly inhibits proliferation in Hep3B cells, which may be related with the activation of ROS/JNK signal pathway.

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更新日期/Last Update: 2019-09-09