三叶青乙酸乙酯提取物对人胰腺癌PANC-1细胞凋亡的影响及机制研究  

Effect of EAF from tetrastigma hemsleyanum root tuber on apoptosis of human pancreatic cancer PANC-1 cells and its mechanism

作　　者：丁小鸽 张春椿[2] 秦海燕[2] 熊耀康[2] 许健[1] 张婷[1,3] 

机构地区：[1]浙江中医药大学医学技术学院,浙江杭州310053 [2]浙江中医药大学药学院 [3]浙江大学生命科学学院

出　　处：《毒理学杂志》2017年第3期182-186,共5页Journal of Toxicology

基　　金：国家中医药行业专项项目(201407002);浙江省科技厅项目(2016C37069)

摘　　要：目的探讨三叶青乙酸乙酯提取物(ethylacetate fraction,EAF)对人胰腺癌PANC-1细胞的凋亡作用及其机制。方法 MTT法检测三叶青EAF对人胰腺癌PANC-1细胞的生长抑制作用;光镜及Hoechst 33258荧光染色分别观察药物作用后细胞形态和细胞凋亡;流式细胞仪检测细胞凋亡率;蛋白印记法(Western blot)检测凋亡蛋白Bax、Bcl-2和P53的表达情况。结果与对照组比较,三叶青EAF能显著地抑制人胰腺癌细胞PANC-1的生长,并呈明显的量-效关系。药物作用后,细胞形态改变,有明显的凋亡特征。流式细胞术检测细胞凋亡率有浓度依赖性。Western blot检测表明三叶青EAF可以促进Bax、P53蛋白表达升高,Bcl-2表达下降。结论三叶青EAF对人胰腺癌PANC-1细胞具有抑制增殖和促进凋亡作用,其机制可能与上调p53和Bax的表达,下调Bcl-2的表达有关。Objective To study the effect of EAF on the proliferation and apoptosis in human pancreatic cancer PANC-1 cells and to explore its possible mechanisms. Methods PANC-1 cells were treated with EAF in different concentrations at different time points. Its inhibition on PANC-1 cell proliferation was detected by MTT assay. The morphology of the PANC-1 cell was observed by inverted microscope. Changes of apoptosis were observed by Hoechst 33258 method. The apoptosis rate was detected by flow cytometry. Expression changes of apoptosis-related proteins Bax,Bcl-2 and P53 were detected by Western blot. Result Compared with the control group,the inhibition rate of PANC-1 cells were increased in a dose and time dependent manner after treated by EAF. Under inverted microscope and fluorescence microscope,typical morphological changes,dark stained nucleus and formed apoptotic body could be seen. Result of flow cytometry showed that the apoptosis rate was increased in a dose-dependent manner treated by EAF. Result of Western blot indicated that compared with the control group,the protein expression of Bax and P53 were up-regulated and that of Bcl-2 were down-regulated treated by different condensations of EAF. Conclusion EAF had obvious effect in inhibiting the PANC-1 cell proliferation and inducing cell apoptosis.The Bax/Bcl2 ratio and P53 were upregulated,which may related with the apoptosis triggered by EAF in PANC-1 cells.

关 键 词：三叶青乙酸乙酯提取物 PANC-1细胞 增殖 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]