中国药理学通报2018，Vol.34Issue(7)：1005-1012,8.DOI:10.3969/j.issn.1001-1978.2018.07.022
miR-519d/Twist1轴介导BrMC抑制SMMC-7721源性肝癌干样细胞体外致癌能力
Inhibition of in vitro carcinogenicity by 8-bromo-7-methoxychrysin mediated by modulating miR-519d/TWIST1 axis in liver cancer stem-like cells from SMMC-7721 cell line
摘要
Abstract
Aim To determine whether 8-bromo-7-me-thoxychrysin ( BrMC) inhibits in vitro carcinogenicity via up-regulating miR-519d expression and down-regu-lating Twist1 expression in liver cancer stem-like cells ( LCSLCs) derived from SMMC-7721 cell line. Meth-ods The second generation spheroids derived from SMMC-7721 cell line were obtained by sphere-forming assay and were considered as LCSLCs . Then LCSLCs were treated with various concentrations ( 1.0, 3.0, 10.0 μmol·L-1) of BrMC. The expression level of miR-519d was detected using real-time PCR. And in vitro carcinogenicity was investigated by sphere-forming assay and clone-forming assay in agar. The transcrip-tional activity and protein expression of Twist1 were an-alyzed using luciferase reporter assay and Western blot. Moreover, the molecular mechanism of BrMC was elucidated via miR-519 mimic transfection and Twist1 gene transduction, respectively. Results Compared with SMMC-7721 cells, miR-519d-3p was low-ex-pressed and Twist1 was over expressed in LCSLCs. And the sphere-forming ratio and the clone-forming ra-tio decreased by treatment with BrMC ( 1.0, 3.0, 10.0 μmol·L-1) in a dose-dependent manner. Fur-thermore, luciferase reporter assay demonstrated miR-519d could directly target the 3′ untranslated region of Twist1 mRNA and regulate protein expression. miR-519d mimic enhanced the effects of BrMC (3.0 μmol ·L-1) . However, Twist1 gene transduction effective-ly reversed the effects of BrMC ( 3.0 μmol·L-1) . Conclusion BrMC inhibits in vivo carcinogenicity via regulating miR-519/Twist1 signal axis in LCSLCs de-rived from SMMC-7721 cell line.关键词
肝细胞癌/肿瘤干细胞/8-溴-7-甲氧基白杨素/治疗作用/miR-519d/Twist1Key words
hepatocellular carcinoma/cancer stem cell/8-bromo-7-methoxychrysin/therapeutic action/miR-519d/Twist1分类
医药卫生引用本文复制引用
罗祎敏,曹晓诚,李翔,崔迎红,许畅,陈阿,曹建国..miR-519d/Twist1轴介导BrMC抑制SMMC-7721源性肝癌干样细胞体外致癌能力[J].中国药理学通报,2018,34(7):1005-1012,8.基金项目
国家自然科学基金资助项目(No 81172375) （No 81172375）
