Identification of valid reference genes for microRNA expression studies in a hepatitis B virus replicating liver cell line

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• 作者：Kari Stougaard Jacobsen ; Kirstine Overgaard Nielsen…
• 关键词：Reference genes ; Normalisation ; microRNA ; Hepatitis B virus ; RT ; qPCR
• 刊名：BMC Research Notes
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：9
• 期：1
• 全文大小：909 KB
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• 作者单位：Kari Stougaard Jacobsen (1) (2)
  Kirstine Overgaard Nielsen (1) (2)
  Thilde Nordmann Winther (1)
  Dieter Glebe (3)
  Flemming Pociot (2)
  Birthe Hogh (1)
  
  1. Department of Paediatrics, Hvidovre Hospital, University of Copenhagen, Copenhagen, Denmark
  2. Department of Paediatrics and Center for Non-Coding RNA in Technology and Health, Herlev Hospital, University of Copenhagen, Copenhagen, Denmark
  3. Institute of Medical Virology, National Reference Center for Hepatitis B and D Viruses, German Center for Infection Research, Biomedical Research Center Seltersberg, Justus-Liebig University Giessen, Giessen, Germany
  
• 刊物主题：Biomedicine general; Medicine/Public Health, general; Life Sciences, general;
• 出版者：BioMed Central
• ISSN：1756-0500

文摘

Background MicroRNAs are regulatory molecules and suggested as non-invasive biomarkers for molecular diagnostics and prognostics. Altered expression levels of specific microRNAs are associated with hepatitis B virus infection and hepatocellular carcinoma. We previously identified differentially expressed microRNAs with liver-specific target genes in plasma from children with chronic hepatitis B. To further understand the biological role of these microRNAs in the pathogenesis of chronic hepatitis B, we have used the human liver cell line HepG2, with and without HBV replication, after transfection of hepatitis B virus expression vectors. RT-qPCR is the preferred method for microRNA studies, and a careful normalisation strategy, verifying the optimal set of reference genes, is decisive for correctly evaluating microRNA expression levels. The aim of this study was to provide valid reference genes for the human HCC-derived cell line HepG2.