光甘草定对肝癌Hep3B细胞增殖的影响  

Effect of glabridin on the proliferation of hepatocarcinoma Hep3B cells

作　　者：周毓旻 沈凌 田烨 孙星[4] ZHOU Yu-min;SHEN Ling;TIAN Ye;SUN Xing(Department of Medical Service,Shanghai First People's Hospital,Shanghai Jiao Tong University,Shanghai 201600;Clinical Medical School,Shanghai First People's Hospital of Nanjing Medical University,Shanghai 201600;Department of Clinical Pharmacy,Shanghai First People's Hospital,Shanghai Jiao Tong University,Shanghai 200080;Department of General Surgery,Shanghai First People's Hospital,Shanghai Jiao Tong University,Shanghai 201600)

机构地区：[1]上海交通大学附属第一人民医院医务处,上海201600 [2]南京医科大学附属上海一院临床医学院,上海201600 [3]上海交通大学附属第一人民医院临床药学科,上海200080 [4]上海交通大学附属第一人民医院普外科,上海201600

出　　处：《中南药学》2020年第9期1483-1487,共5页Central South Pharmacy

基　　金：国家自然科学基金(No.81270556)。

摘　　要：目的探讨光甘草定对肝癌细胞Hep3B增殖的影响。方法运用CCK-8检测光甘草定对肝癌Hep3B细胞活力的抑制作用;流式细胞术检测光甘草定对肝癌Hep3B细胞周期的影响;EDU细胞增殖检测方法探讨光甘草定对肝癌细胞Hep3B增殖的影响;细胞成球实验检测光甘草定对肝癌干细胞成球能力的影响;qRT-PCR法和Western blot法检测光甘草定作用后肝癌Hep3B细胞内相关基因/蛋白表达变化,探讨其作用机制。结果不同浓度光甘草定作用于Hep3B细胞后,细胞活性随着光甘草定浓度的增加而下降,24 h和48 h的IC50分别为7.4μmol·L-1和5.8μmol·L-1。PI染色显示光甘草定导致肝癌Hep3B细胞G2/M期阻滞,EDU细胞增殖实验提示光甘草定可浓度依赖性地抑制Hep3B细胞的增殖;肿瘤干细胞成球实验显示光甘草定显著减弱干细胞成球能力;qRT-PCR法检测发现光甘草定使肿瘤干细胞干性相关基因OCT4、NANOG和SOX2表达降低,同时Western blot法检测发现细胞内Hedgehog信号通路的Gli1蛋白、干性标记物Lgr5和CD133蛋白、c-Myc蛋白表达明显降低。结论光甘草定可能通过Hedgehog信号通路影响肝癌干细胞干性,进而抑制Hep3B细胞增殖。Objective To determine the effect and mechanism of glabridin on the proliferation of the liver cancer Hep3B cells.Methods CCK-8 was used to detect the inhibitory effect of glabridin on Hep3B cell viability.Flow cytometry was used to detect the effect of glabridin on Hep3B cells’cycle.EDU cell proliferation assay was used to determine the proliferation of hepatoma cell Hep3B.Sphere-forming assay was used to detect the effect of glabridin on the sphere formation efficiency of hepatoma stem cells.Western blot and qRT-PCR were used to detect the expression of related genes/proteins in liver cancer Hep3B cells after the effect of glabridin and explore its mechanism.Results The cell activity decreased with the increase of glabridin concentration.The half effective inhibitory concentration(IC50)at 24 h and 48 h was 7.4μmol·L-1 and 5.8μmol·L-1,respectively.PI staining showed that glabridin caused G2/M phase arrest in hepatoma Hep3B cells.EDU cell proliferation experiment suggested that glabridin inhibited Hep3B cells’proliferation in a concentration-dependent manner.Sphere-forming assay of the tumor stem cell showed that glabridin significantly reduced the stem cell sphere-forming abilities;qRT-PCR found that glabridin reduced the expression of stem cell-related genes OCT4,NANOG and SOX2 in the tumor stem cells,while Western blot detected that the expression of Gli1 protein in Hedgehog signaling pathway,stem markers Lgr5,CD133 and c-Myc protein was significantly reduced.Conclusion Glabridin may reduce the stemness of hepatocellular carcinoma stem cells through the Hedgehog signaling pathway,thus inhibiting the proliferation of Hep3B cells.

关 键 词：肝细胞癌 光甘草定 HEDGEHOG信号通路 肿瘤干细胞干性 细胞增殖 

分 类 号：R285[医药卫生—中药学]