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

BMC Research Notes
Kari Stougaard JacobsenBirthe 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 ...Continue Reading

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Citations

Nov 25, 2018·Journal of Cellular Physiology·Kioomars SaliminejadSeyed Hamidollah Ghaffari

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Methods Mentioned

BETA
transfection
PCR
ELISA

Software Mentioned

NormFinder
GraphPad Prism
geNorm

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