ArticleBMC cancer2009
Suitable reference genes for real-time PCR in human HBV-related hepatocellular carcinoma with different clinical prognoses.
Article in BMC cancer, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers, 1 of them a synthesis that pooled it.
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40 citing papers in PubMed, 1 synthesis or guideline pooled it, 64 citations in OpenAlex.
- Identification of reference genes across physiological states for qRT-PCR through microarray meta-analysis.PloS one · 2011Pooled it
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- Transcriptome analysis creates a new era of precision medicine for managing recurrent hepatocellular carcinoma.World journal of gastroenterology · 2023Review
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- Validation of Reference Genes for Normalization of Relative qRT-PCR Studies in Papillary Thyroid Carcinoma.Scientific reports · 2019Article
- Conventionally used reference genes are not outstanding for normalization of gene expression in human cancer research.BMC bioinformatics · 2019Article
- Peroxiredoxin 1, a Novel HBx-Interacting Protein, Interacts with Exosome Component 5 and Negatively Regulates Hepatitis B Virus (HBV) Propagation through Degradation of HBV RNA.Journal of virology · 2019Article
- Tissue Morphology and Gene Expression Characterisation of Transplantable Adenocarcinoma Bearing Mice Exposed to Fluorodeoxyglucose-Conjugated Magnetic Nanoparticles.Journal of biomedical nanotechnology · 2018Article
- Selection of internal references for qRT-PCR assays of human hepatocellular carcinoma cell lines.Bioscience reports · 2017Article
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- Consensus reference gene(s) for gene expression studies in human cancers: end of the tunnel visible?Cellular oncology (Dordrecht, Netherlands) · 2015Review
- Global miRNA expression and correlation with mRNA levels in primary human bone cells.RNA (New York, N.Y.) · 2015Article
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundHousekeeping genes are routinely used as endogenous references to account for experimental differences in gene expression assays. However, recent reports show that they could be de-regulated in different diseases, model animals, or even under varied experimental conditions, which may lead to unreliable results and consequently misinterpretations. This study focused on the selection of suitable reference genes for quantitative PCR in human hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) with different clinical outcomes.
methodsWe evaluated 6 commonly used housekeeping genes' expression levels in 108 HBV-related HCCs' matched tumor and non-tumor tissue samples with different clinical outcomes and 26 normal liver specimens by real-time PCR. The expression stability of the 6 genes was compared using the software programs geNorm and NormFinder. To show the impact of reference genes on data analysis, we took PGK1 as a target gene normalized by each reference gene, and performed one-way ANOVA and the equivalence test.
resultsWith the geNorm and NormFinder software programs, analysis of TBP and HPRT1 showed the best stability in all tissue samples, while 18s and ACTB were less stable. When 18s or ACTB was used for normalization, no significant difference of PGK1 expression (p > 0.05) was found among HCC tissues with and without metastasis, and normal liver specimens; however, dramatically differences (p < 0.001) were observed when either TBP or the combination of TBP and HPRT1 were selected as reference genes.
conclusionTBP and HPRT1 are the most reliable reference genes for q-PCR normalization in HBV-related HCC specimens. However, the well-used ACTB and 18S are not suitable, which actually lead to the misinterpretation of the results in gene expression analysis.
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