阿魏菇乙酸乙酯相三萜类化合物对食管癌Eca109细胞的增殖抑制及机制  

Inhibitory Effect and Mechanism of the Ethyl Acetate Fraction of Pleurotus ferulatus Triterpenoids on Proliferation of Esophageal Eca109 Cells

作　　者：王磊 张富春[1] 刘军[1] WANG Lei;ZHANG Fuchun;LIU Jun(Xinjiang Key Laboratory of Biological Resources and Genetics Engineering,College of Life Scienceand Technology,Xinjiang University,Urumqi 830046,China)

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室

出　　处：《食品科学》2019年第23期189-198,共10页Food Science

基　　金：新疆大学纵向科研项目（61287）;国家自然科学基金青年科学基金项目（31500752）

摘　　要：目的:探讨阿魏菇乙酸乙酯相三萜类化合物(ethyl acetate fraction of Pleurotus ferulatus triterpenoid,PFTP-E)对食管癌Eca109细胞的生长抑制作用及可能机制。方法:采用3-(4,5-二甲基噻-2)-2,5-二苯基四氮唑溴盐法检测PFTP-E体外抑制Eca109细胞的增殖活性;Hoechst 33258染色观察细胞凋亡;流式细胞术检测PFTP-E对Eca109细胞的增殖、周期、凋亡、线粒体膜电位以及胞内活性氧水平变化的影响;免疫印迹法检测PFTP-E对细胞凋亡相关蛋白Bcl2、Bax、聚腺苷二磷酸-核糖聚合酶(poly ADP-ribose polymerase,PARP)、含半胱氨酸的天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase,Caspase)3、Caspase9、细胞周期相关蛋白Cyclin B1、内质网应激相关蛋白真核起始因子2α(eukaryotic initiation factor 2α,eIF2α)、CHOP以及丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路相关蛋白表达的影响。结果:PFTP-E以时间和剂量依赖性抑制Eca109细胞的增殖,并将细胞周期阻滞在G2/M期,进而诱导Eca109细胞凋亡。PFTP-E可引起Eca109细胞线粒体膜电位崩溃并导致胞内活性氧水平升高。PFTP-E可上调细胞色素c、Bax、p-eIF2α、CHOP表达,下调Bcl2、Cyclin B1表达,显著增强剪切型PARP、Caspase3及Caspase9表达,同时诱导内质网应激,激活MAPK/JNK信号通路。结论:PFTP-E能有效诱导Eca109细胞凋亡,其抗肿瘤机制与线粒体损伤途径、周期阻滞、内质网应激等有关。Objective:The study investigated the inhibitory effect of the ethyl acetate fraction of Pleurotus ferulatus triterpenoid(PFTP-E)on the proliferation of human esophageal carcinoma Eca109 cells and the possible underlying mechanism.Methods:Cell proliferation in vitro was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.Hoechst 33258 staining was used to observe cell apoptosis.Flow cytometry was used to assess cell proliferation,cycle,apoptosis,mitochondrial membrane potential and intracellular reactive oxygen species(ROS).The effects of PFTP-E on the expression of the apoptosis-related proteins Bcl2,Bax,poly ADP-ribose polymerase(PARP),caspase 3 and caspase 9;the cell cycle-related protein cyclin B1;the endoplasmic reticulum stress-related proteins eukaryotic initiation factor 2α(eIF2α)and CHOP;and mitogen-activated protein kinase(MAPK)signaling pathway-related proteins were detected by Western blot.Results:PFTP-E inhibited the proliferation of Eca109 cells in both a time and dose-dependent manner,blocked the cell cycle in G2/M phase and induced cell apoptosis.Meanwhile,PFTP-E decreased mitochondrial membrane potential and increased intracellular ROS production.In addition,PFTP-E could up-regulate the expression of cytochrome c,Bax,p-eIF2αand CHOP,down-regulate the expression of Bcl2 and cyclin B1,significantly increase cleaved-PARP,cleaved-caspase 3 and cleaved-caspase 9 expression levels,induce endoplasmic reticulum stress and activate the MAPK/JNK signaling pathway.Conclusion:PFTP-E can effectively induce apoptosis,and the underlying antitumor mechanism is related to mitochondrial damage,cell cycle block,endoplasmic reticulum stress.

关 键 词：阿魏菇 三萜类 ECA109细胞 细胞凋亡 

分 类 号：TS201.4[轻工技术与工程—食品科学]