摘要
[目的]探讨人参皂苷Compound K(人参皂苷CK)通过Bcl-2相关转录因子1(Bclaf1)诱导人肝癌细胞SMMC-7721自噬的作用.[方法]取人参皂苷CK(0,20,40,60,80μmol/L)分别作用于人肝癌细胞SMMC-772124,48,72 h,采用MTT法检测人参皂苷CK对人肝癌细胞SMMC-7721增殖的抑制作用;人参皂苷CK(40μmol/L)处理人肝癌细胞SMMC-772148 h后,单荧光GFP-LC3质粒转染细胞和透射电子显微镜观察敲除Bclaf1基因对细胞自噬的影响,采用Western blot法检测CRISPR/Cas9技术敲除Bclaf1基因后自噬相关蛋白LC3Ⅱ的表达情况.[结果]人参皂苷CK可抑制人肝癌细胞SMMC-7721增殖,与空白对照组比较,Bclaf1表达量明显减少(P<0.01).人参皂苷CK处理Bclaf1稳定敲除肝癌细胞,自噬更加明显,与空白对照组比较,LC3Ⅱ蛋白表达量显著增加(P<0.01).[结论]人参皂苷CK可通过Bclaf1诱导人肝癌细胞SMMC-7721自噬.
OBJECTIVE To explore the effect of ginsenoside Compound K(ginsenoside CK)on autophagy of human hepatoma cell SMMC-7721 induced by bcl-2 related transcription factor 1(Bclaf1).METHODS Ginsenoside CK(0,20,40,60,80μmol/L)was taken to act on human hepatoma cell SMMC-7721 for 24,48 and 72 h respectively,MTT assay was used to detect the inhibition of ginsenoside CK on the proliferation of human hepatoma cell SMMC-7721.After the treatment of human hepatoma cell SMMC-7721 with ginsenoside CK(40μmol/L)for 48 h,single fluorescent GFP-LC3 transfection and transmission electron microscopy assays were used to observe the effect of Bclaf1 knockout on autophagy.Western blot was used to detect autophagy related protein LC3Ⅱexpression after the knockout of Bclaf1 gene with CRISPR/Cas9.RESULTS Ginsenoside CK could inhibit the proliferation of human hepatoma cell SMMC-7721.Compared with the control group,Bclaf1 expression was significantly reduced(P<0.01).Autophagy was more obvious on the Bclaf1 stable knockout hepatoma cell after the treatment of ginsenoside CK.Compared with the control group,the expression of LC3Ⅱprotein increased significantly(P<0.01).CONCLUSION Ginsenoside CK could induce autophagy in human hepatoma cell SMMC-7721 through Bclaf1.
作者
陈佳欣
闫岩
苏杰琳
赵嘉琪
张学武
CHEN Jiaxin;YAN Yan;SU Jielin;ZHAO Jiaqi;ZHANG Xuewu(Department of Biochemistry and Molecular Biology,Yanbian University College of Medicine,Yanji 133002,Jilin,China)
出处
《延边大学医学学报》
CAS
2020年第3期157-161,共5页
Journal of Medical Science Yanbian University
基金
国家自然科学基金(81760728)。
