摘要
目的肺癌位居我国居民癌症发病率和死亡率的首位,其中非小细胞癌(non-small cell lung cancer,NSCLC)约占80%,筛选高效低毒的抗癌药物尤为迫切。本研究拟探讨姜黄素对NSCLC细胞的可能作用机制。方法用不同浓度的姜黄素(0、10、20、30μmol/L)或活性氧清除剂(CAT和NAC)加姜黄素处理肺癌细胞A549和SPC-A1,采用流式细胞术检测细胞周期、细胞凋亡率、活性氧(reactive oxygen species,ROS)水平和线粒体膜电位的变化,蛋白质印迹法检测自噬相关蛋白LC3、P62和细胞凋亡相关蛋白PARP、Caspase-3和Caspase-9的表达变化。结果姜黄素抑制非小细胞肺癌细胞株A549和SPC-A1增殖及克隆形成,主要将细胞阻滞在G_2/M期,0、10、20和30μmol/L姜黄素处理的A549细胞G_2/M期细胞百分比分别为(12.67±2.52)%、(22.67±2.52)%、(27.00±2.01)%和(42.33±4.04)%,SPC-A1细胞G_2/M期细胞百分比分别为(9.33±2.52)%、(18.33±1.53)%、(20.67±2.52)%和(30.67±1.53)%。0、10、20和30μmol/L姜黄素处理A549细胞凋亡率分别为(4.40±1.02)%、(7.31±1.52)%、(9.32±1.08)%和(13.97±1.98)%,P<0.05;SPC-A1细胞凋亡率分别为(4.38±1.22)%、(5.98±0.75)%、(9.42±1.25)%和(16.13±3.09)%,P<0.05。姜黄素导致ROS水平增高、线粒体膜电位降低和线粒体自噬的发生,并且呈剂量依赖性,而应用ROS清除剂可以减弱以上药物作用。相关信号转导通路蛋白表达与以上细胞生物行为改变表现一致。结论姜黄素通过ROS途径诱导NSCLC细胞发生线粒体自噬,是一种有潜力的抗癌药物。
OBJECTIVE In recent years,lung cancer is the most commonly diagnosed cancer and the leading cause of cancer death for both men and women in China and screening high efficiency and low toxicity of anti-cancer drugs is particularly urgent.Therefore,we intended to explore the possible mechanism of curcumin on non-small cell lung cancer（NSCLC）in this study.METHODS NSCLC cells were treated with different concentrations of curcumin（0,10,20,30μmol/L）or oxidative stress scavenger（CAT,NAC）plus curcumin.MTS,clone formation experiment,flow cytometry and Western Blot were involved in investigating the effect of curcumin on tumor cell proliferation,cell cycle,apoptosis,ROS level,JC-1and related signal transduction pathways.RESULTS In vitro,curcumin inhibited the proliferation of NSCLC cell lines and mainly induced G_2/M phase arrest.The data are as follows：the percentage of G_2/M phase cells in A549 cells was（12.67±2.52）%,（22.67±2.52）%,（27.00±2.01）%and（42.33±4.04）%,respectively.The ratio of G_2/M phase cells in each group of SPC-A1were（9.33±2.52）%,（18.33±1.53）%,（20.67±2.52）%and（30.67±1.53）%（P〈0.05）.Besids,curcumin induced cell apoptosis and the apoptotic rates of A549 cells were（4.40±1.02）%,（7.31±1.52）%,（9.32±1.08）% and（13.97±1.98）%,respectively,in each treatment group（DMSO,10,20,30μmol/L）.The apoptotic rates of SPC-A1 treated group were（4.38±1.22）%,（5.98±0.75）%,（9.42±1.25）% and（16.13±3.09）%,respectively（P〈0.05）.Curcumin also resulted in ROS accumulation,mitochondrial membrane potential decreased in a dose-dependent manner,and the application of oxidative stress scavenger can reduce the anti-cancer effect of curcumin.The expression of related signal transduction pathway protein was consistent with the above biological behavior.CONCLUSION Curcumin induces NSCLC cells apoptosis via the reactive oxygen species-mediated mitochondrial autophagy pathway.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2017年第10期663-669,共7页
Chinese Journal of Cancer Prevention and Treatment
基金
山东省医药卫生科技发展计划(2015WS0167)
关键词
姜黄素
活性氧簇
线粒体自噬
抗癌作用
curcumin
EF24
reactive oxygen species
mitochondrial autophagy
anti-cancer effect
