吉马酮通过HBXIP/p53信号通路诱导A549、CNE-1、HepG2细胞凋亡  

Apoptosis of A549,CNE-1,and HepG2 cells induced by gematone through HBXIP/p53 signal pathway

作　　者：刘建军[1] 赵营莉 唐海涛 代文婷[1] 夏泉[3] 方兴[1] LIU Jianjun;ZHAO Yingli;TANG Haitao;DAI Wenting;XIA Quan;FANG Xing(The Second People's Hospital of Hefei,Hefei 230000,China;Anhui International Travel Healthcare Center(Hefei Customs Port Clinic),Hefei 230000,China;The First Affiliated Hospital of Anhui Medical University,Hefei 230000,China)

机构地区：[1]合肥市第二人民医院(安徽医科大学附属合肥医院)广德路药学部,安徽合肥230000 [2]安徽国际旅行卫生保健中心(合肥海关口岸门诊部),安徽合肥230000 [3]安徽医科大学第一附属医院,安徽合肥230000

出　　处：《药物评价研究》2023年第5期1024-1031,共8页Drug Evaluation Research

基　　金：合肥市自然科学基金(2021012);安徽省高等学校科学研究项目(自然科学类)(2022AH050712);合肥市第二人民医院院级课题(2021ygkt35)。

摘　　要：目的研究吉马酮诱导肺癌A549、鼻咽癌CNE-1、肝癌HepG2细胞凋亡的机制。方法50、150、200、250、300μmol·L^(−1)的吉马酮处理肝癌HepG2、肺癌A549、鼻咽癌CNE-1、结肠癌Caco-2细胞24、48、72 h后,MTT实验检测细胞的存活率的变化。100、150、200μmol·L^(−1)的吉马酮分别处理A549、HepG2、CNE-1细胞48 h后采用流式细胞术检测细胞凋亡的变化;Western blotting实验检测细胞凋亡标志蛋白cleaved-Caspase-3、Caspase-3的变化,检测乙型肝炎X相互作用蛋白(HBXIP)、p53蛋白的表达量变化。分别在A549、HepG2、CNE-1细胞中应用siRNA敲低HBXIP,Western blotting实验检测HBXIP的敲低效果及p53蛋白表达变化;MTT实验检测敲低HBXIP对吉马酮诱导的细胞增殖抑制作用的影响。结果吉马酮对鼻咽癌CNE-1、肝癌HepG2细胞增殖抑制作用较强,与溶剂对照组比较,200、250、300μmol·L^(−1)组细胞存活率显著下降(P<0.05);在高浓度时对肺癌A549细胞增殖抑制效果较强,与溶剂对照组比较,250、300μmol·L^(−1)组细胞存活率显著下降(P<0.05);对结肠癌Caco-2细胞作用相对较弱。与对照组比较,100、150、200μmol·L^(−1)吉马酮处理A549、HepG2、CNE-1细胞48 h后,凋亡率显著升高(P<0.05),cleaved-Caspase-3、p53蛋白表达显著上升(P<0.05);以150、200μmol·L^(−1)吉马酮处理A549、HepG2、CNE-1细胞48 h后,HBXIP的蛋白表达显著降低(P<0.05)。siRNA-HBXIP处理48 h后,与对照组比较,HBXIP的蛋白表达量显著下降(P<0.05),p53蛋白表达量显著上升(P<0.05)。与单独使用的相同浓度的吉马酮相比,沉默HBXIP后A549、HepG2、CNE-1细胞对吉马酮的敏感度明显提高,150、200、250μmol·L^(−1)组均差异显著(P<0.05)。结论吉马酮可以抑制A549、HepG2、CNE-1细胞增殖、诱导细胞凋亡,作用机制可能与调控HBXIP/p53信号通路相关。Objective Study the mechanism of gemmazone induced apoptosis in A549,CNE-1,and HepG2 cells.Methods After 50,150,200,250,and 300μmol·L^(−1) germacrone treated in HepG2,A549,CNE-1,and Caco-2 cells for 24,48 and 72 h,the MTT assay was preformed to detect the changes of cell viability.When 100,150 and 200μmol·L^(−1) germacrone treated in A549,HepG2 and CNE-1 cells for 48 h,the changes of cell apoptosis were detected by flow cytometry,and the Western blotting was used to detect the changes of cleaved-Caspase 3,Caspase-3 HBXIP,and P53 proteins.The expressions of HBXIP and P53 were detected by Western blotting after HBXIP knocked down in A549,HepG2 and CNE-1 cells.The effect of knockdown of HBXIP on the inhibitory proliferation induced by germacrone was detected by MTT assay.Results Gemmazone had a strong inhibitory effect on the proliferation of nasopharyngeal carcinoma CNE-1 and liver cancer HepG2 cells,compared with the solvent control group,the cell survival rate of the 200,250,and 300μmol·L^(−1) group significantly decreased(P<0.05).At high concentrations,the inhibitory effect on the proliferation of lung cancer A549 cells was better,compared with the solvent control group,the cell survival rate of the 250,300μmol·L^(−1) group significantly decreased(P<0.05).The effect on colon cancer Caco-2 cells was relatively weak.Compared with the control group,after 48 hours of treatment with 100,150,200μmol·L^(−1) gemmazone,the apoptosis rate of A549,HepG2,and CNE-1 cells significantly increased(P<0.05),and the expression of cleaved-Caspase-3 and p53 proteins significantly increased(P<0.05).After 48 hours of treatment with 150,200μmol·L^(−1) gemmazone in A549,HepG2,and CNE-1 cells,the protein expression of HBXIP was significantly reduced(P<0.05).After 48 hours of siRNA-HBXIP treatment,compared with the control group,the protein expression of HBXIP significantly decreased(P<0.05)and the expression of p53 protein significantly increased(P<0.05).Compared with the same concentration of gemmazone used alone

关 键 词：吉马酮 肝癌 肺癌 鼻咽癌 结肠癌 A549细胞 CNE-1细胞 HEPG2细胞 增殖 凋亡 HBXIP p53 

分 类 号：R285.5[医药卫生—中药学]