摘要
目的研究薯蓣皂苷(DIO)对人HepG2肝癌细胞凋亡的影响及其可能抗癌作用机制。方法DIO(0.25、0.5、1、2、4、6和8μmol/L)干预HepG2肝癌细胞,MTT法检测细胞增殖情况,并计算半数抑制率(IC50);划痕实验分析细胞的迁移能力,Western blot检查细胞中凋亡因子及Wnt/β-catenin通路蛋白表达差异。结果与对照组比较,DIO组HepG2细胞抑制率显著升高(P<0.01),并诱导细胞在形态学上出现凋亡特点,呈时间与剂量依赖性;划痕实验结果显示DIO显著抑制HepG2细胞的迁移距离;与对照组比较,DIO干预细胞24 h后,Caspase-3、cleaved Caspase-3水平明显上调,而Bcl-2、β-catenin水平显著下调(P<0.05、P<0.01);采用LiCl激活Wnt/β-catenin信号通路,LiCl组细胞中Wnt1、β-catenin水平较对照组显著上调(P<0.01);与LiCl组比较,LiCl+DIO组HepG2细胞Wnt1、β-catenin、GSK-3β蛋白表达显著下调(P<0.01)。结论DIO体外能够促进HepG2肝癌细胞凋亡,机制可能是其下调β-catenin的蛋白表达,抑制Wnt/β-catenin信号通路传导,进而抑制凋亡因子Bcl-2表达,最终诱导细胞凋亡而发挥抗肝癌作用。
Objective To detect the apoptosis effects of dioscin in HepG2 cells and its possible anti-hepatocellular carcinoma mechanisms.Methods HepG2 human hepatocellular carcinoma cells were exposed to 0.25,0.5,1,2,4,6,or 8μmol/L dioscin,and cell proliferation was measured via MTT assay.The half-maximal inhibitory concentration(IC50)was calculated with the software.A scratch test was used to analyze cell migration ability.Western blot was employed to evaluate the expression of apoptosis and Wnt/β-catenin-pathway-related proteins.Results Compared with the control group,the dioscin-treated HepG2 cells’proliferation was significantly more inhibited,and the inhibition increased in a time-and dose-dependent manner(P<0.01).HepG2 cells showed morphological characteristics of apoptosis after they were treated with 1μmol/L or 2μmol/L dioscin.The scratch test indicated that the migration distance of HepG2 cells was remarkably reduced when treated with dioscin.In the Western blot experiment,the expression levels of Caspase-3 and cleaved Caspase-3 were visibility up-regulated,while those of Bcl-2 andβ-catenin were significantly down-regulated when the cells were treated with dioscin for 24 h(P<0.05,P<0.01).When LiCl reagent was added to the HepG cells to activate the Wnt/β-catenin signaling pathway,the expression levels of Wnt1 andβ-catenin were remarkably increased compared with those of the control group(P<0.01).Compared with the LiCl group,the LiCl+DIO group’s expression of Wnt1,β-catenin,and GSK-3βwas significantly decreased(P<0.01).Conclusions DIO can promote the apoptosis of HepG2 cells by inhibitingβ-catenin protein expression and thereby down-regulating the Wnt/β-catenin signaling pathway.This inhibits apoptosis-related gene Bcl-2 expression,which leads to the induction of cell apoptosis.Therefore,DIO can have an anti-hepatocellular carcinoma effect.
作者
梁玉琼
黄庆
黄娟娟
梁芳
滕丽娟
郑洋
LIANG Yuqiong;HUANG Qing;HUANG Juanjuan;LIANG Fang;TENG Lijuan;ZHENG Yang(Experiment Teaching Center,Faculty of Chinese Medicine Science Guangxi University of Chinese Medicine,Nanning 530222,China;Department of Pharmacy,the First Affiliated Hospital of Guangxi University ofTraditional Chinese Medicine,Nanning 530023)
出处
《中国比较医学杂志》
CAS
北大核心
2024年第8期72-77,86,共7页
Chinese Journal of Comparative Medicine
基金
2022年度广西高校中青年教师科研基础能力提升项目(2022KY1671)
广西自然科学基金项目(2023GXNSFBA026230)
广西中医药大学赛恩斯新医药学院科研项目(2023MS004)。
关键词
薯蓣皂苷
HEPG2
肝癌
WNT/Β-CATENIN信号通路
细胞凋亡
dioscin
HepG2
hepatocellular carcinoma
Wnt/β-catenin signaling pathway
apoptosis Conflicts of Interest:The authors declare no conflict of interest
