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
10.3969/j.issn.1001-1978.2018.07.022
- VernacularTitle: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
- Author:
Yi-Min LUO
1
;
Xiao-Cheng CAO
;
Xiang LI
;
Ying-Hong CUI
;
Chang XU
;
A CHEN
;
Jian-Guo CAO
Author Information
1. 南华大学医学院病理学教研室
- Keywords:
hepatocellular carcinoma;
cancer stem cell;
8-bromo-7-methoxychrysin;
therapeutic action;
miR-519d;
Twist1
- From:
Chinese Pharmacological Bulletin
2018;34(7):1005-1012
- CountryChina
- Language:Chinese
-
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.
