ArticleBMC research notes2016
Identification of valid reference genes for microRNA expression studies in a hepatitis B virus replicating liver cell line.
Article in BMC research notes, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 17 citations in OpenAlex.
- Unlocking the Future of Hepatocellular Carcinoma Early Diagnosis: The Promise of Extracellular Vesicle Biomarkers.Journal of clinical and translational hepatology · 2026Review
- miR-128-3p and miR-223-3p as Potential Biomarkers of Metabolic Dysfunction in Liver Tissue from Patients Undergoing Metabolic Bariatric Surgery.Obesity surgery · 2026Article
- Analysis of qRT-PCR Data to Identify the Most Stable Reference Gene Using gQuant.Bio-protocol · 2025Article
- Development of a robust and generalizable algorithm "gQuant" for accurate normalizer gene selection in qRT-PCR analysis.Scientific reports · 2024Article
- Identification of reference microRNAs in skeletal muscle of a canine model of Duchenne muscular dystrophy.Wellcome open research · 2024Article
- miRNA Expression Profiling in Subcutaneous Adipose Tissue of Monozygotic Twins Discordant for HIV Infection: Validation of Differentially Expressed miRNA and Bioinformatic Analysis.International journal of molecular sciences · 2022Article
- Reference Genes for qPCR-Based miRNA Expression Profiling in 14 Human Tissues.Medical principles and practice : international journal of the Kuwait University, Health Science Centre · 2022Review
- Selection of reference genes for quantitative analysis of microRNA expression in three different types of cancer.PloS one · 2022Article
- Reference genes identification for normalization of qPCR under multiple stresses inPlant methods · 2018Article
- Selection of Reliable Reference Genes for Gene Expression Studies onFrontiers in plant science · 2016Article
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Authors and funding
6 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundMicroRNAs 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.
resultsA 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.
conclusionWe 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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