The effect of miR-155 on HBV replication and PTEN expression in vivo
10.3760/cma.j.issn.1007-3418.2018.07.002
- VernacularTitle: miR-155对乙型肝炎病毒复制及PTEN表达的影响
- Author:
Cong XIE
1
;
Guangli REN
2
;
Mancun XU
2
;
Weiyun ZHANG
3
;
Sulin ZHANG
3
;
Qiyin CAI
2
;
Yongmin LIN
2
;
Donglong ZHOU
Author Information
1. Department of Pediatric, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010, China (Department of Pediatric, the Secon Affiliated Hospital of Guangzhou Medical University)
2. Department of Pediatric, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010, China
3. Department of Clinical Laboratory, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010, China
- Publication Type:Journal Article
- Keywords:
Hepatitis B virus;
Interferon-gamma;
phosphatase and tensin homology deleted on chromosome ten;
micro RNA155
- From:
Chinese Journal of Hepatology
2018;26(7):489-494
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the mmu-miR-155 eukaryotic overexpression vector pmR-155 and to investigate its effect on HBV replication and expression of PTEN in vivo.
Methods:The mmu-mir-146a precursor gene fragment pre-mmu-mir-146a was amplified by PCR, then connected to the pmR-mCherry plasmid vector after double enzyme digestion, the accuracy of recombinant vector was verified by colony PCR、double enzyme digestion and sequencing; then the recombinant vector was transfected HBV transgene mice(Experimental Group)with hydrodynamics-based injection via vena caudalis, and pmR-mCherry plasmid、PBS were respectively transfected into the mice as Empty plasmid Group、Blank Group. The concentration of IFN-γ in the serum was detected by ELISA. The expression of SOCS1、PTEN mRNA in the liver was detected by qPCR at 30d post-transfectioned. The Western blot was performed to detect the changes in SOCS1、PTEN、HBX in the liver tissue at 30 d post-transfectioned. The results were analyzed with Student’s t-test, or one-way analysis of variance and the least significant difference test.
Results:the colony PCR、double enzyme digestion and sequencing verified that the gene was inserted into the pmR-mCherry vector. Compared with Blank Group, the expression of miR-155 in the Experimental Group was significantly increased(t = 8.90, P < 0.01); the concentration of IFN-γ in the Experimental Group was significantly increased(F = 26.58, P < 0.01); the mRNA(FSOCS1 mRNA = 19.72, P < 0.01; FPTEN mRNA = 7.38, P < 0.05) and protein(FSOCS1 = 50.30, P < 0.01; FPTEN = 129.00, P < 0.01) expression of COCS1、PTEN was significantly decreased in the Experimental group and the protein of HBX was also significantly(FHBX = 77.97, P < 0.01).
Conclusion:The pmR-155 eukaryotic overexpression vector is successfully constructed, this recombinant vector can express miR-155 stably; miR-155 can down-regulate cocs1、PTEN gene expression and up-regulate the expression of IFN-γ, it can inhibit the replication of HBV and a potential targets to treating hepatocellular carcinoma.
