摘要
目的研究吴茱萸碱对鼻咽癌5-8F细胞增殖、凋亡及PI3K/AKT信号通路的影响。方法(1)将5-8F细胞分为溶剂对照组、不同浓度吴茱萸碱组(0.5、1.0、2.0、4.0μmol·L^(-1))、顺铂组(cisplatin,4.0μg·mL^(-1));(2)将5-8F细胞设为5组:溶剂对照组、SC792.5μmol·L^(-1)组、SC79+吴茱萸碱2.0μmol·L^(-1)组、吴茱萸碱2.0μmol·L^(-1)组、LY29400250μmol·L^(-1)组。采用实时无标记细胞功能分析仪(real time cellular analysis technology,RTCA)监测细胞增殖的情况;Annexin V-FITC/PI双荧光染色法检测细胞凋亡率,Hoechest 33342染色法观察细胞凋亡形态;Western blot法检测磷酸肌醇3-激酶蛋白(phosphatidylinositol 3-kiases,PI3K)、磷酸化蛋白质激酶蛋白B(phosphorylated protein kinase B,p-AKT)、增殖细胞核抗原蛋白(proliferating cell nuclear antigen,PCNA)、X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis,XIAP)、存活蛋白(Survivin)的表达水平。结果与溶剂对照组相比,不同浓度吴茱萸碱组(0.5、1.0、2.0、4.0μmol·L^(-1))显著抑制5-8F细胞增殖(P<0.05或P<0.01);吴茱萸碱组(1.0、2.0μmol·L^(-1))的细胞核荧光染色增强,凋亡率明显升高(P<0.01);PI3K、p-AKT、PCNA、XIAP、Survivin蛋白表达水平明显下降(P<0.05或P<0.01)。与吴茱萸碱2.0μmol·L^(-1)组比较,SC79+吴茱萸碱2.0μmol·L^(-1)组的PI3K、p-AKT、PCNA、XIAP、Survivin蛋白表达水平升高(P<0.05或P<0.01),且吴茱萸碱抑制5-8F细胞增殖和诱导凋亡的效应降低(P<0.05或P<0.01)。结论吴茱萸碱可能通过抑制PI3K/AKT信号通路活性,下调增殖及凋亡相关蛋白PCNA、XIAP、Survivin的表达水平,最终抑制鼻咽癌细胞增殖和诱导凋亡。
Objective To investigate the effects of evodiamine on proliferation and apoptosis of 5-8F cells in nasopharyngeal carcinoma cells and the role of PI3K/AKT signaling pathway.Methods The 5-8F cells were divided into solvent control group,evodiamine(0.5,1.0,2.0,4.0μmol·L^(-1))groups and cisplatin(4.0μgmol·L^(-1))group;in addition,the 5-8F cells were divided into 5 groups:solvent control group,SC79(2.5μmol·L^(-1))group,SC79+evodiamine(2.0μmol·L^(-1))group,evodiamine(2.0μmol·L^(-1))group,and LY294002(50μmol·L^(-1))group.The cell proliferation was monitored by real time cellular analysis technology(RTCA),the apoptosis rate was detected by annexin V-FITC/PI double fluorescence staining,and the apoptosis morphology was observed by Hoechest 33342 staining.The expression levels of phosphatidylinositol 3-kinase(PI3K),phosphorylated protein kinase B(p-AKT),proliferating cell nuclear antigen(PCNA),X-linked inhibitor of apoptosis(XIAP)and Survivin were detected by Western blot.Results Compared with the solvent control group,different concentrations of evodiamine(0.5,1.0,2.0,4.0μmol·L^(-1))significantly inhibited the proliferation of 5-8F cells(P<0.05 or P<0.01),and the nuclear fluorescence staining and apoptosis rate of cells in evodiamine group(1.0,2.0μmol·L^(-1))increased significantly(P<0.01).The protein expression of PI3K,p-AKT,PCNA,XIAP and Survivin decreased significantly(P<0.05 or P<0.01).Compared with evodiamine(2.0μmol·L^(-1))group,the protein expression levels of PI3K,p-AKT,PCNA,XIAP and Survivin in SC79+evodiamine(2.0μmol·L^(-1))group were elevated(P<0.05 or P<0.01),and the effects of evodiamine on inhibiting proliferation and inducing apoptosis of 5-8F cells decreased(P<0.05 or P<0.01).Conclusion Evodiamine may inhibit proliferation and induce apoptosis of nasopharyngeal carcinoma cells by inhibiting the activity of PI3K/AKT signaling pathway and down-regulating the expression of proliferation and apoptosis-related proteins PCNA,XIAP and Survivin.
作者
郭利培
刘洁
张文青
史红健
范婧莹
王贤文
何迎春
GUO Lipei;LIU Jie;ZHANG Wenqing;SHI Hongjian;FAN Jingying;WANG Xianwen;HE Yingchun(Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Hunan Provincial Key Laboratory for the Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine,Changsha,Hunan 410208,China;Hunan Provincial Ophthalmology and Otolaryngology Diseases Prevention and Treatment with Chinese Medicine and Visual Function Protection Engineering and Technological Research Center,Changsha,Hunan 410208,China;The First Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410007,China)
出处
《湖南中医药大学学报》
CAS
2023年第4期612-618,共7页
Journal of Hunan University of Chinese Medicine
基金
国家自然科学基金项目(81973914,81874408)
湖南省教育厅项目(21B0358,21C0241)
湖南省中医药管理局项目(D2022105)
2021年度湖南中医药大学校级科研基金项目(2021XJJJ014,2021XJJJ008)
湖南省卫生健康委员会项目(D202307017740)。
关键词
鼻咽癌细胞
吴茱萸碱
增殖
凋亡
PI3K/AKT信号通路
nasopharyngeal carcinoma cell
evodiamine
proliferation
apoptosis
PI3K/AKT signaling pathway
