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Overexpression of DNA-methyltransferases in persistency of cccDNA pool in chronic hepatitis B

Костюшев Д. С., Зуева А. П., Брезгин С. А., Липатников А. Д., Симирскии В. Н., Глебе Д., Волчкова Е. В., Шипулин Г. А., Чуланов В. П.
Терапевтический архив
Т. 89, Вып. 11, С. 21-26
Опубликовано: 2017
Тип ресурса: Статья

DOI:10.17116/terarkh2017891121-26

Аннотация:
Aim. To define the role of DNA-methyltransferases of type 1 and type 3A in hepatitis B viral cycle. Materials and methods. Human hepatoma cells HepG2 with stable expression of 1.1-mer HBV genome were transfected with vectors encoding DNA-methyltransferase 1 (DNMT1), DNA-methyltransferase 3A (DNMT3A) or were co-transfected with these vectors. Total HBV DNA copy number, relative expression of pregenomic RNA (pgRNA), S-protein-encoding RNA (S-RNA) and cccDNA were analyzed by quantitative and semi-quantitative real-time PCR-analysis with TaqMan probes for assessment of DNMTs-mediated effects on HBV. Results. DNMT1 and DNMT3A suppress HBV transcription and replication, though to different magnitude. cccDNA pool is enlarged statistically significantly 2-fold (P<0.005) after transfection of DNMT3A, but is unaltered under DNMT1 treatment. Conclusion. DNMT3A regulates the size of cccDNA pool and is important for persistency of HBV infection.
Ключевые слова:
CccDNA; Chronic hepatitis B (CHB); DNA-methyltransferases (DNMT); Hepatitis B virus (HBV); PCR; PgRNA
circular DNA; DNA (cytosine 5) methyltransferase; DNA (cytosine 5) methyltransferase 1; DNA methyltransferase 3A; DNMT1 protein, human; chronic hepatitis B; Hep-G2 cell line; Hepatitis B virus; human; metabolism; DNA (Cytosine-5-)-Methyltransferase 1; DNA (Cytosine-5-)-Methyltransferases; DNA, Circular; Hep G2 Cells; Hepatitis B virus; Hepatitis B, Chronic; Humans
Язык текста: Русский
ISSN: 2309-5342
Костюшев Д. С.
Зуева А. П.
Брезгин С. А.
Липатников А. Д.
Симирскии В. Н.
Глебе Д.
Волчкова Е. В. Елена Васильевна 1953-
Шипулин Г. А.
Чуланов В. П. Владимир Петрович 1970-
Kostyushev D. S.
Zueva A. P.
Brezgin S. A.
Lipatnikov A. D.
Simirskii V. N.
Glebe D.
Volchkova E. V. Elena Vasilyevna 1953-
Shipulin G. A.
Chulanov V. P. Vladimir Petrovich 1970-
Overexpression of DNA-methyltransferases in persistency of cccDNA pool in chronic hepatitis B
Текст визуальный непосредственный
Терапевтический архив
Медицинское маркетинговое агентство "МедиаМедика"
Т. 89, Вып. 11 С. 21-26
2017
Статья
CccDNA Chronic hepatitis B (CHB) DNA-methyltransferases (DNMT) Hepatitis B virus (HBV) PCR PgRNA
circular DNA DNA (cytosine 5) methyltransferase DNA (cytosine 5) methyltransferase 1 DNA methyltransferase 3A DNMT1 protein, human chronic hepatitis B Hep-G2 cell line Hepatitis B virus human metabolism DNA (Cytosine-5-)-Methyltransferase 1 DNA (Cytosine-5-)-Methyltransferases DNA, Circular Hep G2 Cells Hepatitis B virus Hepatitis B, Chronic Humans
Aim. To define the role of DNA-methyltransferases of type 1 and type 3A in hepatitis B viral cycle. Materials and methods. Human hepatoma cells HepG2 with stable expression of 1.1-mer HBV genome were transfected with vectors encoding DNA-methyltransferase 1 (DNMT1), DNA-methyltransferase 3A (DNMT3A) or were co-transfected with these vectors. Total HBV DNA copy number, relative expression of pregenomic RNA (pgRNA), S-protein-encoding RNA (S-RNA) and cccDNA were analyzed by quantitative and semi-quantitative real-time PCR-analysis with TaqMan probes for assessment of DNMTs-mediated effects on HBV. Results. DNMT1 and DNMT3A suppress HBV transcription and replication, though to different magnitude. cccDNA pool is enlarged statistically significantly 2-fold (P<0.005) after transfection of DNMT3A, but is unaltered under DNMT1 treatment. Conclusion. DNMT3A regulates the size of cccDNA pool and is important for persistency of HBV infection.