10-羟基癸烯酸诱导肝癌细胞凋亡、周期阻滞及迁移抑制的机制  

Mechanism by which 10-Hydroxy-2-decenoic Acid Induces Cell Apoptosis and Cycle Arrest and Inhibits Migration in Hepatocellular Carcinoma HepG2 Cells

作　　者：臧延青[1] 鞠雪莹 翟雨晴 姚笛[1] 朱磊[1] 王长远[1,2] ZANG Yanqing;JU Xueying;ZHAI Yuqing;YAO Di;ZHU Lei;WANG Changyuan(College of Food Science,Heilongjiang Bayi Agricultural University,Daqing 163319,China;National Coarse Cereals Engineering Research Center,Daqing 163319,China)

机构地区：[1]黑龙江八一农垦大学食品学院,黑龙江大庆163319 [2]国家杂粮工程技术研究中心,黑龙江大庆163319

出　　处：《食品科学》2021年第23期182-195,共14页Food Science

基　　金：“十三五”国家重点研发计划重点专项(SQ2018YFE020070);黑龙江省自然科学基金项目(QC2017024);黑龙江省青年创新人才培养项目(UNPYSCT-2017108);黑龙江八一农垦大学研究生创新科研资助项目(YJSCX2019-Y51)。

摘　　要：10-羟基癸烯酸(10-hydroxy-2-decenoic acid,10-HDA)是蜂王浆中特有的活性成分,本实验以肝癌HepG2细胞为研究对象,通过细胞毒性实验(cell counting kit-8,CCK-8)、Hoechst 33342/碘化丙啶(propidium iodide,PI)双染、流式细胞术及蛋白免疫印迹(Western blot)法分析10-HDA预处理后HepG2细胞凋亡的分子机制。结果发现,10-HDA对肝癌细胞系有明显的细胞毒性,对正常肝、肺、胃细胞(L-02、IMR-90、GES-1)无明显作用。同时,HepG2细胞经10-HDA处理后,Bcl-2表达水平下调、Bax表达水平上调,触发了线粒体凋亡途径,线粒体膜电位下降,从而导致细胞色素c(cytochrome c,Cyt c)得到释放,Caspase-3被活化,最终线粒体依赖性凋亡。此外,10-HDA能够诱导活性氧(reactive oxygen species,ROS)累积,激活了丝裂原活化蛋白激酶(mitogenactivated protein kinase,MAPK)和信号转导与转录活化因子(signal transducer and activator of transcription,STAT)3信号通路,促进P-Jun氨基末端激酶(jun N-terminal kinase,JNK)、P-P38表达,抑制P-细胞外调节蛋白激酶(extracellular-signal-regulated kinase,ERK)、P-STAT3表达,进而调控细胞周期相关蛋白,使细胞周期阻滞在G2/M期。最后,10-HDA通过激活转化生长因子-β1(transforming growth factor-β1,TGF-β1)信号通路,验证了其抑制迁移作用。综上所述,10-HDA通过累积ROS、激活MAPK和STAT3信号通路,诱导细胞凋亡。10-Hydroxy-2-decenoic acid(10-HDA)is a unique bioactive component of royal jelly.In this study,the molecular mechanism of apoptosis in HepG2 cells induced by 10-HDA was analyzed by cell counting kit-8(CCK-8)assay,Hoechst 33342/propidium iodide(PI)staining,flow cytometry and Western blot.The results showed that 10-HDA had significant cytotoxicity on liver cancer cell lines but not normal cells(L-02,IMR-90 or GES-1).10-HDA down-regulated the expression of Bcl-2,up-regulated the expression of Bax,triggered the mitochondrial apoptosis pathway,and decreased mitochondrial membrane potential,thus leading to the release of cytochrom c(Cyt c),the activation of caspase-3,and eventually mitochondria-dependent apoptosis.In addition,10-HDA induced reactive oxygen species(ROS)accumulation,activated the mitogen-activated protein kinase(MAPK)signaling pathway and the signal transducer and activator of transcription 3(STAT3)signaling pathway,promoted the expression of jun N-terminal kinase(JNK)and P-P38,and inhibited the expression of extracellular-signal-regulated kinase(ERK)and P-STAT3,consequently regulating the expression of cell cycle related-proteins to arrest the cell cycle at the G2/M phase.Finally,10-HDA was demonstrated to inhibit cancer cell migration by activating the transforming growth factor-β1(TGF-β1)signaling pathway.In conclusion,10-HDA can induce cell apoptosis by causing ROS accumulation and activating the MAPK and STAT3 signaling pathways.

关 键 词：10-羟基癸烯酸 肝癌细胞 细胞凋亡 周期阻滞 细胞迁移 信号通路 

分 类 号：R735.7[医药卫生—肿瘤]