miR-200c targets nuclear factor IA to suppress HBV replication and gene expression via repressing HBV Enhancer I activity

• Published in: Biomedicine & pharmacotherapy 2018-03, Vol.99, p.774-780
• Main Authors: Tian, Hui, He, Zhenkun
• Format: Article
• Language: English
• Subjects: Blotting, Western; Cell Line, Tumor; DNA, Viral - metabolism; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation, Viral; Genes, Reporter; HBV Enhancer I; Hep G2 Cells; Hepatitis B e Antigens - metabolism; Hepatitis B Surface Antigens - metabolism; Hepatitis B virus; Hepatitis B virus - physiology; Hepatitis B, Chronic - genetics; Hepatitis B, Chronic - virology; Humans; Luciferases - genetics; MicroRNAs - genetics; miR-200c; NFI Transcription Factors - genetics; NFIA; replication; RNA, Viral - metabolism; Virus Replication - physiology
• ISSN: 0753-3322; 1950-6007
• DOI: 10.1016/j.biopha.2018.01.141

Hepatitis B virus (HBV) chronic infection is a health problem in the worldwide, with a underlying higher risk of liver cirrhosis and hepaticocellular carcinoma. A number of studies indicate that microRNAs (miRNAs) play vital roles in HBV replication. This study was designed to explore the potential molecular mechanism of miR-200c in HBV replication. The expression of miR-200c, nuclear factor IA (NFIA) mRNA, HBV DNA, and HBV RNA (pregenomic RNA (pgRNA), and total RNA) were measured by qRCR. The levels of HBsAg and HBeAg were detected by ELISA. NFIA expression at protein level was measured by western blot. The direct interaction between miR-200c and NFIA were identified by Targetscan software and Dual-Luciferase reporter analysis. Enhance I activity were detected by Dual-Luciferase reporter assay. miR-200c expression was prominently reduced in pHBV1.3-tranfected Huh7 and in stable HBV-producing cell line (HepG2.2.15). The enforced expression of miR-200c significantly suppressed HBV replication, as demonstrated by the reduced levels of HBV protein (HBsAg and HBeAg) and, DNA and RNA (pgRNA and total RNA) levels. NFIA was proved to be a target of miR-200c and NFIA overexpression notably stimulated HBV replication. In addition, the inhibitory effect of miR-200c on HBV Enhance I activity was abolished following restoration of NFIA. miR-200c repressed HBV replication by directly targeting NFIA, which might provide a novel therapeutic target for HBV infection.