Effects and mechanism of miR-214 on hepatocellular carcinoma
10.1016/S1995-7645(14)60350-3
- Author:
Li-Li ZHANG
1
;
Yan-Jun GUO
1
;
Chun-Na ZHAO
1
;
Jian-Yun GAO
1
Author Information
1. Department of Gastroenterology, Qiqihaer Medical College Affiliated training hospital Daqing Oil Field General Hospital
- Publication Type:Journal Article
- Keywords:
Hepatocellular carcinoma;
MiR-214;
β-catenin pathway
- From:
Asian Pacific Journal of Tropical Medicine
2015;8(5):392-398
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To explore the role of miR-214 in the progression of hepatocellular carcinoma (HCC) and its inhibitory mechanisms in depressing the signaling pathway of β-catenin, this study was conducted. Methods: We ectopically expressed miR-214 in HepG2 cells to obtain cell lines Lv-miR-214-HepG2 and their control Lv-control-HepG2. Differences between the two cell lines were compared in cell growth, proliferation, colony forming ability and cell cycles. RT-PCR method was applied for the quantification of β-catenin mRNA expression. Western-blot method was applied for the determination of the protein level of β-catenin and their downstream targets (ie. Cyclin D1, c-Myc and TCF-1). The effect of miR-214 on cells was further explored through RNA interference and restoring miR-214 expression. Results: In comparison with negative (Lv-control-HepG2) and blank (HepG2) control, a significant inhibition of cell growth and proliferation caused by miR-214 was observed after 48~72h of cell culture experiments (P<0.05). The miR-214 treatment resulted in a colony forming efficiency of (23.28±3.26)%, which was significantly lower than that of negative control [(51.31±3.97)%] (P<0.05). According to FCM results, the experimental group, compared with control, showed a higher proportion of cells in G0/G1 phase [(70.32±3.12)%] but a lower proportion in S phase [(18.42±2.90)%] (P<0.05). The MTT assay demonstrated a significant inhibition of the proliferation and β-catenin expression of HCC cells compared with control (P<0.05), while no significant difference was observed after HCC cells being transfected with β-catenin overexpression plasmid (P>0.05). By comparing to the RT-PCR and Western-blot results of control, the miR-214 treatment led to a slightly decrease in the β-catenin mRNA expression (P>0.05), but an extremely inhibition in the protein level of β-catenin and its downstream targets Cyclin D1, c-Myc, and TCF-1 (P<0.05). Conclusions: miR-214 functions as a suppressor during the progression of HCC, and its inhibitory role was achieved by down-regulating β-catenin signaling pathway.
