α-常春藤皂苷通过激活和稳定p53/Noxa信号诱导肝癌细胞凋亡  

α-Hederin Induces Apoptosis in Hepato-cellular Carcinoma Cells by Activating and Stabilizing p53/Noxa Signaling Pathway

作　　者：陈晓静 周莉 刘凯琪 段菊凤 刘明 李洪亮 汪选斌 CHEN Xiaojing;ZHOU Li;LIU Kaiqi;DUAN Jufeng;LIU Ming;LI Hongliang;WANG Xuanbin(Department of Pharmacy of Renmin Hospital and School of Pharmaceutical Sciences,Hubei University of Medicine,Shiyan 442000,China;Biomedical Research Institute,Hubei University of Medicine,Shiyan 442000,China;Hubei Key Laboratory of Wudang Local Chinese Medicine Research,Hubei University of Medicine,Shiyan 442000,China)

机构地区：[1]湖北医药学院附属人民医院药学部和药学院,十堰442000 [2]湖北医药学院生物医药研究院,十堰442000 [3]武当特色中药研究湖北省重点实验室(湖北医药学院),十堰442000

出　　处：《医药导报》2024年第3期334-345,共12页Herald of Medicine

基　　金：国家自然科学基金面上项目(82274155);湖北省科技厅对外合作项目(2022EHB046);湖北省科技厅自然基金面上项目(2023AFB925);武当特色中药研究湖北省重点实验室开放课题(WDCM2018002,WDCM201917,WDCM201918);“十四五”湖北省高等学校优势特色学科群(现代医学)项目(2022XKQY3)。

摘　　要：目的研究预知子活性成分α-常春藤皂苷对肝癌细胞的体内外抑制作用及其机制。方法将肝癌细胞分为4组,分别给予0、10、20和30μmol·L^(-1)α-常春藤皂苷至24、48 h。噻唑蓝(MTT)法检测细胞增殖,流式细胞术检测凋亡率,转录组学筛选α-常春藤皂苷作用的信号通路,核糖核酸(RNA)干扰、实时荧光定量聚合酶链式反应(qRT-PCR)和蛋白免疫印迹检测信号通路和凋亡通路信使核糖核酸(mRNA)和蛋白表达,体内移植瘤试验验证α-常春藤皂苷体内对肝癌细胞生长的抑制作用。结果α-常春藤皂苷通过激活腺苷二磷酸核糖聚合酶(PARP)、天冬氨酸特异性半胱氨酸蛋白酶9(Caspase9)和Caspase3诱导肝癌细胞凋亡。转录组学、qRT-PCR和蛋白免疫印迹检测发现α-常春藤皂苷上调了p53和Noxa的表达,并抑制p53和Noxa蛋白的降解,α-常春藤皂苷还能逆转p53/Noxa敲低所致的凋亡减少。动物实验结果表明,α-常春藤皂苷可抑制肝癌细胞移植瘤的增殖。结论α-常春藤皂苷可通过激活和稳定p53/Noxa信号诱导肝癌细胞凋亡。Objective To investigate the inhibitory effects and mechanisms ofα-hederin,an active ingredient in Fructus Akebiae,on hepatocellular carcinoma(HCC)cells.Methods HCC cells were divided into four groups and treated withα-hederin(0,10,20,and 30μmol·L^(-1))for 24 h and 48 h,respectively.MTT assays were used to detect the cell proliferation rate,flow cytometry(FCM)was used to detect the apoptotic rate,transcriptomics was used to screen signaling pathways inα-hederin-treated HCC cells,RNA interference was exploited to verify the underlying signaling pathway,and real-time quantitative PCR(qRT-PCR)and Western blotting(WB)were used to detect expression changes of the mRNA and protein of TP53(p53),PMAIP1(Noxa),and apoptosis-associated proteins,Caspase9 and Caspase3.Resultsα-Hederin induced apoptosis by activating apoptosis-associated proteins,PARP,Caspase9 and Caspase3.Transcriptomics,qRT-PCR,and WB results also showed thatα-hederin increased the mRNA and protein expression of p53 and Noxa.Furthermore,α-hederin inhibited the protein degradation of p53 and Noxa,reversing the apoptosis decrease in p53/Noxa siRNA-knocked-down HCC cells.In vivo results showed thatα-hederin inhibited the growth of HCC tumors.Conclusionα-hederin may induce the apoptosis of HCC cells by activating and stabilizing the p53/Noxa signaling pathway.

关 键 词：α-常春藤皂苷 肝细胞癌 凋亡 P53 NOXA 

分 类 号：R286[医药卫生—中药学] R965[医药卫生—中医学]