Terapevticheskii arkhivTerapevticheskii arkhivТерапевтический архив0040-36602309-5342LLC Obyedinennaya Redaktsiya3228710.17116/terarkh2017891121-26Editorial articleПередовая статьяResearch ArticleOverexpression of DNA-methyltransferases in persistency of cccDNA pool in chronic hepatitis BРоль гиперэкспрессии ДНК-метилтрансфераз в персистенции кольцевой ковалентно замкнутой ДНК при хроническом гепатите ВKostyushevD SКостюшевД СZuevaA PЗуеваА ПBrezginS AБрезгинС АLipatnikovA DЛипатниковА ДSimirskiiV NСимирскийВ НGlebeDVolchkovaE VВолчковаЕ ВShipulinG AШипулинГ АChulanovV PЧулановВ ПФБУН «Центральный научно-исследовательский институт эпидемиологии» РоспотребнадзораФГБОУ ВО «Московский государственный университет им. М.В. Ломоносова»ФГАОУ ВО «Первый МГМУ им. И.М. Сеченова» Минздрава РоссииФГБОУ ВО «Российский химико-технологический университет им. Д.И. Менделеева» Минобрнауки РоссииФГБУН «Институт биологии развития им. Н.К. Кольцова» РАНГессенский университет им. Ю. ЛибихаФГАОУ ВО «Первый МГМУ им. И.М. Сеченова» Минздрава РоссииФБУН «Центральный научно-исследовательский институт эпидемиологии» Роспотребнадзора151120178911VOL 89, NO11 (2017)ТОМ 89, №11 (2017)212610042020Copyright ©; 2017, Consilium MedicumCopyright ©; 2017, ООО "Консилиум Медикум"2017Consilium MedicumООО "Консилиум Медикум"https://creativecommons.org/licenses/by-nc-sa/4.0https://ter-arkhiv.ru/0040-3660/article/view/32287

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.

Резюме Цель исследования. Изучение роли ДНК-метилтрансфераз 1-го и 3А типов в жизненном цикле вируса гепатита В (HBV). Материалы и методы. Клетки гепатомы человека HepG2, стабильно экспрессирующие геном HBV 1.1-mer, трансфицированы векторами, которые кодируют ДНК-метилтрансферазу-1 (ДНМТ1), ДНК-метилтрансферазу-3А (ДНМТ3А) или котрансфицированы обоими векторами. Определение общего числа копий ДНК HBV, уровней экспрессии прегеномной РНК (pgRNA), РНК, кодирующей S-белок (S-RNA), и уровень кольцевой ковалентно замкнутой ДНК (ккзДНК) методом количественного и полуколичественного анализа c помощью полимеразной цепной реакции в реальном времени с зондами TaqMan использовали для оценки действия ДНК-метилтрансфераз на жизненный цикл HBV. Результаты. ДНМТ1 и ДНМТ3А в разной степени подавляют транскрипцию и репликацию HBV. Количество ккзДНК статистически значимо увеличивается под действием ДНМТ3А, но не изменяется под действием ДНМТ1. Заключение. ДНМТ3А регулирует количество ккзДНК и является важным фактором персистенции HBV.

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