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

Overview of attention for article published in BMC Research Notes, January 2016
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Title
Identification of valid reference genes for microRNA expression studies in a hepatitis B virus replicating liver cell line
Published in
BMC Research Notes, January 2016
DOI 10.1186/s13104-016-1848-2
Pubmed ID
Authors

Kari Stougaard Jacobsen, Kirstine Overgaard Nielsen, Thilde Nordmann Winther, Dieter Glebe, Flemming Pociot, Birthe Hogh

Abstract

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. A panel of 739 microRNAs was screened to identify the most stably expressed microRNAs, followed by a PubMed search identifying microRNAs previously used as reference genes. Sixteen candidate reference genes were validated by RT-qPCR. Reference gene stabilities were calculated first by standard deviations of ΔCt values and then by geNorm and NormFinder analyses, taking into account the amplification efficiency of each microRNA primer set. The optimal set of reference genes was verified by a target analysis using RT-qPCR on miR-215-5p. We identified miR-24-3p, miR-151a-5p, and miR-425-5p as the most valid combination of reference genes for microRNA RT-qPCR studies in our hepatitis B virus replicating HepG2 cell model.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 25 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
China 1 4%
Unknown 24 96%

Demographic breakdown

Readers by professional status Count As %
Researcher 5 20%
Other 5 20%
Student > Bachelor 3 12%
Student > Ph. D. Student 3 12%
Student > Master 2 8%
Other 4 16%
Unknown 3 12%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 10 40%
Medicine and Dentistry 5 20%
Agricultural and Biological Sciences 3 12%
Pharmacology, Toxicology and Pharmaceutical Science 1 4%
Veterinary Science and Veterinary Medicine 1 4%
Other 2 8%
Unknown 3 12%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 24 January 2016.
All research outputs
#18,436,183
of 22,840,638 outputs
Outputs from BMC Research Notes
#3,017
of 4,266 outputs
Outputs of similar age
#285,815
of 395,188 outputs
Outputs of similar age from BMC Research Notes
#101
of 139 outputs
Altmetric has tracked 22,840,638 research outputs across all sources so far. This one is in the 11th percentile – i.e., 11% of other outputs scored the same or lower than it.
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