β-榄香烯通过抑制HIF-1α表达促进人肺癌细胞凋亡

• 作 者: 付有政 温华
• 机 构: 中国医科大学附属第一医院呼吸内科,沈阳110001;本溪市中心医院呼吸内科,本溪117000;中国医科大学附属第一医院呼吸内科,沈阳,110001
• 出 处: 解剖科学进展
• 发表日期: 2017年6期
• 关键词: Apoptosis A549 cells β-elemene HIF-1α PI3K/Akt signaling pathway

目的 探讨β-榄香烯是否通过抑制肺癌细胞中HIF-1 α的高表达促进肺癌细胞的凋亡及相关机制.方法 常规培养人肺癌A549细胞,实验分为对照组、乏氧组、β-榄香烯组、不同浓度β-榄香烯/乏氧处理组;通过转染含有HIF-1 α基因的质粒过表达HIF-1 α基因.应用AnnexinV-FITC凋亡检测试剂盒及流式细胞仪、TUNEL检测细胞凋亡.并采用免疫荧光检测HIF-1α蛋白的活化、Westen blot方法检测Bax、caspase-3及Bcl-xL蛋白表达及转染效率.结果 β-榄香烯能够抑制肺癌A549细胞的增殖力,呈时间和浓度依赖性,能够诱导乏氧状态下的肺癌A549细胞凋亡.β-榄香烯可通过上调A549细胞的Bax、caspase-3蛋白表达,降低Bcl-xL表达来诱导乏氧状态下肺癌A549的凋亡.此外,β-榄香烯可有效降低乏氧状态下HIF-1 α蛋白的高表达.通过转染过表达HIF-1α基因,大大降低了β-榄香烯的诱导凋亡作用.结论 β-榄香烯可以上调乏氧状态下A549细胞凋亡,可能与抑制HIF-1 α的高表达相关.“,”Objective To investigate the effects of β-Elemene on apoptosis of human lung cancer A549 cells via inhibition of HIF-1 α protein expression.Methods A549 cells were cultured and supplemented with 10％ FBS,1％ penicillin/streptomycin in 5％ CO2 at 37° C.The cells were classified into four groups:control group,20μ g/m1 β-Elemene group,hypoxia group,hypoxia/10 μ g/ml β-Elemene group and hypoxia/20 μ g/ml β-Elemene group.The cell apoptosis was assessed by flow cytometry using the Annexin V-FITC/propidium iodide assay kit or TENEL to differentiate the survival and apoptotic cells.Bax,Bcl-xL,caspase-3 and HIF-1 α proteins expressions were detected by Western blot or immunofluorescence staining.Results β-Elemene inhibited the proliferation of A549 cells in a dose-time dependent manner,induced the apoptosis of the cells under hypoxia,which might be related to the upregulation of Bax and caspase-3 expressions and downregulation of Bcl-xL expression.In addition,β-Elemene decreased effectively the overexpression of HIF-1 α under hypoxia,but the effect could be downregulated by overexpression of HIF-1 α in A549 cells.Conclusion The increased A549 cells apoptosis under hypoxia condition by β-Elemene might be related to the inhibition of HIF-1α protein expression.