文章摘要
何蕾,王宝辉,吕望,泮辉,胡坚.藤黄酸诱导肺癌细胞H1975凋亡的机制研究[J].,2015,37(13):1124-1128.
藤黄酸诱导肺癌细胞H1975凋亡的机制研究
Gambogic acid induces apoptosis of lung cancer H1975 cells by activating JNK signaling pathway and upregulating ROS
  
DOI:
中文关键词: 藤黄酸  肺癌H1975细胞  活性氧自由基  JNK 信号通路  凋亡
英文关键词: Gambogic acid  Lung cancer H1975 cell  Reactive oxygen species  JNK signaling pathway  Apoptosis
基金项目:浙江中医药大学校级课题
作者单位
何蕾 浙江大学附属第一医院心胸外科 
王宝辉 浙江中医药大学附属第一医院中心实验室 
吕望 浙江大学附属第一医院心胸外科 
泮辉 浙江大学附属第一医院心胸外科 
胡坚 浙江大学附属第一医院心胸外科 
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中文摘要:
      目的 研究藤黄酸(GA)诱导人肺癌细胞H1975凋亡的分子机制,探讨氧自由基(ROS)和JNK信号通路在GA杀伤 肺癌细胞中的作用。方法 以人肺癌细胞H1975为研究对象,MTT法测定GA抑制细胞增殖的作用,Annexin V/PI 双染法测定细胞凋亡率,DCFH-DA 法测定ROS 含量,JC-1探针染色分析线粒体膜电位(MMP),Western blot检测JNK 信号通路的激活和线粒体凋亡途径相关蛋白表达的变化。结果 GA 呈剂量依赖性抑制H1975细胞的增殖,各实验组细胞存活率与空白对照组比较,均有统计学差异(P<0.05 或0.01)。1、2.5 和5滋mol/L GA 作用24h 后,细胞凋亡率分别为25.2%、51.8%和75.1%,剪切型凋亡相关蛋白cleaved caspase-9、cleaved caspase-3 和cleaved PARP 的表达随GA 浓度增高而显著增加,与空白对照组比较,均有统计学差异(P<0.05或0.01)。GA 作用2h 后H1975细胞ROS 含量显著升高,磷酸化JNK(p-JNK)表达上调(P<0.05或0.01)。GA 作用16h后各实验组细胞MMP 均明显降低(均P<0.05)。GA 作用24h后实验组细胞线粒体凋亡途径相关蛋白Bax、Bak、Bik表达增加,而抗凋亡蛋白Bcl-2表达与空白对照组相比明显下降(P<0.05 或0.01)。结论 GA 具有诱导H1975 细胞凋亡的作用,其可能机制是上调细胞内ROS含量,激活JNK 信号通路,进而引起MMP 降低和线粒体凋亡途径激活。
英文摘要:
      Objective To investigate the effect of gambogic acid (GA) on apoptosis of human lung cancer H1975 cells and its mechanisms. and JNK signaling pathway in killing lung cancer cells by GA. Methods Human lung cancer H1975 cells were treated with GA (1, 2.5 and 5滋mol/L). Cell proliferation was analyzed by MTT assay; cell apoptosis was detected by Annexin V-FITC/PI double staining. The Reactive oxygen species (ROS) was measured by DCFH-DA method; the changes of mitochondrial membrane potential (MMP) were detected by JC-1 probe method; the activation of JNK signaling pathway and the expression of mitochondrial apoptosis-related proteins were detected by Western blot. Results Compared with control group, the proliferation of H1975 cells was inhibited by GA in a dose-dependent manner (P<0.05 or 0.01): the apoptotic rates were increased by 25.2%, 51.8% and 75.1%, after cells were treated with 1, 2.5 and 5滋mol/L of GA for 24h. The expression of cleaved caspase-9, cleaved caspase-3 and cleaved PARP in GA group were significantly higher than those in control group in a dose-dependent manner (P<0.05 or 0.01). Compared with control group, the ROS levels were significantly increased and the expression of phospho-JNK(p-JNK) was up-regulated after treated with GA for 2h. After treated by GA for 16 h, the MMP in GA group was decreased significantly compared with the control group(P<0.05). GA increased the expression of pro-apoptotic protein Bax, Bak, Bik and decreased the expression of pro-survival protein Bcl-2 (P<0.05 or 0.01).Conclusion Gambogic acid can induce the apoptosis of human lung cancer H1975 cells through up-regulating the intracellular level of ROS to activate JNK signaling pathway, the latter triggers loss of MMP and activation of mitochondria apoptosis pathways.
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