新藤黄酸通过Ras/Raf/Erk信号通路诱导肺腺癌A549细胞凋亡的研究  

Effect of Gambogenic Acid on Inducing A549 Cell Apoptosis Through Regulating Ras/Raf/Erk Signaling Pathway

作　　者：彭程[1] 孙美灵 苏婧婧[3,4] 李庆林[3,4] 

机构地区：[1]安徽中医药大学,安徽合肥230012 [2]中国科学院成都生物研究所,四川成都610041 [3]安徽中医药大学科研实验中心,安徽合肥230012 [4]新安医学教育部重点实验室,安徽合肥230012

出　　处：《中药新药与临床药理》2016年第2期189-193,共5页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然基金面上项目(81173600);安徽中医药大学自然科学研究基金项目(2014zr023)

摘　　要：目的研究新藤黄酸(Gambogenic acid,GNA)诱导A549细胞凋亡的作用,并探讨其可能的分子机制。方法采用噻唑蓝(MTT)法检测0.5,1.0,2.0,4.0,8.0μmol·L^(-1)的GNA作用于A549细胞24 h后细胞的增殖情况;采用Hoechst 33342和Annexin V-FITC/PI染色法,于荧光显微镜下观察1.25,2.5,5.0μmol·L^(-1)的GNA作用细胞24 h后的凋亡形态;流式细胞仪检测1.25,2.5,5.0μmol·L^(-1)的GNA作用细胞24 h后的凋亡率;Western Blot法检测1.25,2.5,5.0μmol·L^(-1)的GNA处理细胞24 h后,Ras、Raf、Erk和p-Erk的表达。结果GNA能够显著降低A549细胞的活力,并呈浓度依赖性抑制细胞增殖,24 h的IC50为2.507μmol·L^(-1);Hoechst33342和AV/PI染色后,于荧光显微镜下观察发现,细胞显示明显的凋亡特征;流式细胞仪检测结果表明,随着GNA浓度的升高,细胞的凋亡比率逐渐上升;Western Blot法检测显示GNA能够抑制Ras、Raf、p-Erk1/2蛋白的表达。结论 GNA能够诱导A549细胞凋亡,其作用可能与通过调控Ras/Raf/Erk信号通路有关。Objective To investigate the effect of gambogenic acid（GNA） on inducing apoptosis of A549 cells,and to explore its possible molecular mechanism. Methods Methyl thiazolyl tetrazolium（MTT）assay to detect the activity of A549 cell after being treated with 0.5, 1.0, 2.0, 4.0, 8.0 μmol/L of GNA for 24 h. We observed the apoptotic morphology of A549 cells treated with 1.25, 2.5, 5.0 μmol/L of GNA for 24 h under fluorescence microscope after Hoechst 33342 and Annexin V- FITC/PI fluorescence staining. Flow cytometry assay was applied to detect the apoptotic rate of A549 cells after being treated with 1.25, 2.5, 5.0 μmol/L of GNA for 24 h. Western blot assay was used to detect Ras, Raf, Erk and p- Erk expression in A549 cells treated with 1.25, 2.5, 5.0 μmol/L of GNA for 24 h.Results GNA significantly reduced A549 cell activity in a dose- dependent manner and the IC50 with 24 h was2.507μmol/L. Obvious apoptotic characteristics of the cells were shown under fluorescence microscope after Hoechst33342 and AV/PI staining. Flow cytometry assay results showed that the apoptotic rate was increased with the concentration of GNA,and GNA also inhibited the protein expression of Ras,Raf,p- Erk1/2. Conclusion GNA can induce the apoptosis of A549 cells through Ras/Raf /Erk signaling pathway.

关 键 词：新藤黄酸 A549细胞 凋亡 Ras/Raf/Erk信号通路 肺癌 

分 类 号：R285.5[医药卫生—中药学]