Aldo-keto reductase family 1 B10 participates in the regulation of hepatoma cell cycle through p27/p-Rb signaling pathway
10.3760/cma.j.cn501113-20191113-00418
- VernacularTitle:醛酮还原酶家族1成员B10通过p27/p-Rb信号通路参与肝癌细胞周期调控
- Author:
Nan GENG
1
;
Yuanyuan JIN
;
Shixuan ZHU
;
Yurong LI
;
Leyu ZHENG
;
Weijia ZHU
;
Yanwei LI
;
Chao HAN
;
Xiaoguang DOU
;
Han BAI
Author Information
1. 中国医科大学附属盛京医院感染科，沈阳 110022
- Keywords:
Hepatocellular carcinoma;
Aldo keto reductase1B10;
Epalrestat;
shRNA;
Cell cycle arrest
- From:
Chinese Journal of Hepatology
2020;28(10):861-867
- CountryChina
- Language:Chinese
-
Abstract:
Objective:Aldo-keto reductase family 1 member B10 (AKR1B10) pathogenesis, early diagnosis and prognosis are closely related with hepatoma. Therefore, this study explores the effect and mechanism of AKR1B10 on cell cycle in hepatoma cells.Methods:HepG2 cells were infected with lentivirus LV-AKR1B10-shRNA or treated with epalrestat, an AKR1B10 inhibitor. The expression level of AKR1B10 was detected by Western blot assay and real-time fluorescence quantitative PCR (RT-qPCR). Decreased AKR1B10 activity was detected by reduced coenzyme II (NADPH) absorbance at 340 nm. The low expression of AKR1B10 and the effect of different concentrations of epalrestat on cell proliferation and cell cycle were detected by CCK-8 method and flow cytometry. The protein expression levels of p-rb, cyclin D1, E1, p27 in HepG2 cells were detected by Western blot. The mean of the two samples was tested using independent sample t-test.Results:AKR1B10 expression level in hepatoma cells was significantly increased compared to normal liver cells, and the relative expression level of AKR1B10 protein in HepG2 cells was 6.71 ± 1.11 ( P = 0.012). Epalrestat was significantly inhibited with the enzymatic activity of AKR1B10 in a dose-dependent manner. AKR1B10 gene in HepG2 cells was effectively silenced. HepG2 cells treated with different concentrations of epalrestat (AKR1B10 inhibitor) for 24, 48 and 72 h had inhibited cell proliferation, promoted G0/G1 cell cycle arrest, reduced the expression of p-Rb, cyclin D1, and cyclin E1 and increased the expression of cyclin dependent kinase inhibitor p27 expression. Conclusion:AKR1B10 inhibitory expression and activity can promote G0/G1 cell cycle arrest in HepG2 cells through the p27 / p-Rb pathway.
