白花丹醌抑制肝癌类干细胞增殖迁移及干性转录因子的表达  

Inhibition of proliferation and migration of hepatocellular carcinoma⁃like stem cells and expression of stemness transcription factors by Plumbagin

作　　者：周凤玲 陈婉君 黄德伦 马远征 袁彬 叶一娴 韦晓洁 韦燕飞 ZHOU Fengling;CHEN Wanjun;HUANG Delun;MA Yuanzheng;YUAN Bin;YE Yixian;WEI Xiaojie;WEI Yanfei(Guangxi University of Traditional Chinese Medicine,School of Basic Medical Sciences,Nanning 530200,China)

机构地区：[1]广西中医药大学基础医学院,广西南宁530200

出　　处：《海南医学院学报》2024年第14期1059-1067,共9页Journal of Hainan Medical University

基　　金：广西自然科学基金重点项目(2019GXNSFDA245003);广西一流学科建设开放课题(2019XK140);广西中医药大学博士科研启动基金项目(2018BS012,2020BS012);广西中医药大学第三批“岐黄工程”高层次人才团队培育项目(202402)。

摘　　要：目的:探索白花丹醌(Plumbagin, PLB)对肝癌类干细胞增殖迁移及干性转录因子表达的影响。方法:采用多细胞肿瘤球体模式培养富集肝癌类干细胞,设置肝癌细胞组与肝癌类干细胞组,PLB浓度0、5、7.5、10μmol/L组,药物作用时间为24 h,采用MTT法、平板克隆实验、悬浮干细胞成球实验和划痕实验检测细胞增殖、体外成球和迁移能力;qRTPCR和Western blotting检测干性基因和干性蛋白的表达。结果:成功富集肝癌HepG2类干细胞,与肝癌HepG2细胞组比较,肝癌HepG2类干细胞组细胞克隆形成率降低、划痕愈合率和体外成球率升高,差异具有统计学意义(P<0.05);与PLB 0μmol/L组比较,PLB 5、7.5、10μmol/L组,肝癌HepG2类干细胞克隆形成率、体外成球率和划痕愈合率均降低,EpCAM、BMI1、cMYC、Oct4、Nanog干性相关基因及Notch信号通路mRNA的表达水平降低,cMYC蛋白表达水平降低,差异具有统计学意义(P<0.05)。结论:PLB可抑制肝癌HepG2类干细胞增殖、迁移及体外成球能力,从而抑制肝癌细胞干性,其机制可能与下调EpCAM、BMI1、cMYC、Oct4、Nanog干性相关基因和Notch信号通路mRNA的表达水平,以及cMYC蛋白的表达水平有关。Objective:To investigate the effects of plumbagin(PLB)on the proliferation,migration and expression of stem-ness transcription factors of hepatocellular carcinoma stem cells.Methods:The liver cancer stem cells were enriched by multi-cell tumor sphere culture mode.The liver cancer cell group and liver cancer stem cell group were set up with PLB concentration 0,5,7.5,10μmol/L groups,and drug treatment time was 24 h.The cell proliferation,sphere formation and migration ability in vitro were detected by MTT assay,plate cloning assay,suspension stem cell sphere formation assay and scratch test.The expression of stemness genes and proteins was detected by qRT-PCR and Western blotting.Results:HepG2 stem cells were successfully en-riched.Compared with HepG2 cell group,the clone formation rate of HepG2 stem cells group was decreased,and the scratch heal-ing rate and sphere formation rate in vitro were increased,and the differences were statistically significant(P<0.05).Compared with the PLB 0μmol/L group,the colony formation rate,sphere formation rate and scratch healing rate of HepG2 stem cells in the PLB 5,7.5,10μmol/L groups were decreased.The mRNA expression levels of EpCAM,BMI1,c-MYC,Oct-4,Nanog stemness related genes and Notch signaling pathway were decreased,and the protein expression level of c-MYC was decreased(P<0.05).Conclusion:PLB can inhibit the proliferation,migration and sphere formation ability of hepatoma-like stem cells in vitro,thereby inhibiting the stemness of hepatoma cells.The mechanism may be related to the down-regulation of the mRNA expression levels of EpCAM,BMI1,c-MYC,Oct-4,Nanog stemness related genes and Notch signaling pathway,and the expression level of c-MYC protein.

关 键 词：白花丹醌 肝癌细胞干性 干性标记物 增殖 迁移 

分 类 号：R285[医药卫生—中药学]