二氯乙酸盐增强盐酸吡柔比星杀伤肝癌细胞的研究  

Dichloroacetate enhances cytotoxicity of pirarubicin in hepatocellular carcinoma cells in vitro

作　　者：闫小晶[1] 倪振洪[1] 吴亚冉[1] 梁书龙 秦利燕[3] 何凤田[1] 连继勤[1] 

机构地区：[1]第三军医大学基础医学部生物化学与分子生物学教研室,重庆400038 [2]第三军医大学学员旅11营,重庆400038 [3]第三军医大学西南医院血液科,重庆400038

出　　处：《第三军医大学学报》2016年第6期570-574,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金项目(81572375);重庆市自然科学基金项目(CSTC2014jcyjA10110)~~

摘　　要：目的探讨二氯乙酸盐(Dichloroacetate,DCA)增强盐酸吡柔比星(Pirarubicin,THP)杀伤肝癌细胞的效果及可能机制。方法用40μmol/L DCA与不同浓度(0、200、400、600、800μmol/L)盐酸吡柔比星(THP)联合处理肝癌Hep G2细胞24 h,采用CCK-8法检测对细胞增殖的影响。然后用40μmol/L二氯乙酸盐与600μmol/L盐酸吡柔比星联合处理肝癌Hep G2细胞24 h,用Western blot检测细胞中PARP的表达变化及JNK的磷酸化水平;用DCFH-DA荧光探针检测细胞内ROS的水平。结果 DCA可显著增强不同浓度THP对肝癌Hep G2细胞的杀伤效果,与PBS组相比具有统计学差异(P<0.05);DCA联合THP处理24 h可显著升高细胞内的ROS水平,与PBS联合THP组相比具有统计学差异(P<0.05);此外,DCA联合THP可使Hep G2细胞中凋亡相关分子PARP的剪切明显增加,并增强JNK的磷酸化水平。抗氧化剂NAC可显著降低DCA与THP联合处理对Hep G2细胞中JNK的激活作用。结论 DCA可显著增强THP杀伤肝癌细胞效应,其机制可能与ROS/JNK通路的激活有关。Objective To investigate the synergistic effect of dichloroacetate （DCA） and pirarubicin （THP） in hepatocellular carcinoma （HCC） cells and the possible mechanism. Methods HepG2 cells were treated with DCA （40 μmol/L） combined with THP with different concentrations （0, 200, 400, 600, and 800 μmol/L） for 24 h, and then CCK-8 assay was used to detect the cell viability. Moreover, the cells were treated with or without 600 μmol/L of THP in the presence or absence of 40 μmol/L of DCA. Then Western blotting was utilized to measure the expression of cellular molecules, and DCFH-DA fluorescent probes were used to estimate the level of reactive oxygen species （ROS）. Results Compared with PBS, DCA significantly enhanced the cytotoxicity of THP in HepG2 cells （P〈0.05）. The combination of DCA and THP induced an obvious increase of ROS compared with the combination of PBS and THP （P〈0.05）. Moreover, the combination of DCA and THP obviously increased the cleavage of PARP and the phosphorylation of JNK. Antioxidant NAC could significantly inhibit JNK phosphorylation in HepG2 cells treated with DCA and THP. Conclusion DCA significantly enhances the cytotoxicity of THP in HCC cells, which may be dependent on the activation of ROS/JNK signaling pathway.

关 键 词：二氯乙酸盐 肝癌 盐酸吡柔比星 活性氧簇 C-JUN氨基末端激酶 

分 类 号：R73-361[医药卫生—肿瘤] R735.7[医药卫生—临床医学]