ArticleBMC molecular biology2008
PSMB2 and RPL32 are suitable denominators to normalize gene expression profiles in bronchoalveolar cells.
Article in BMC molecular biology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed, 67 citations in OpenAlex.
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- Investigation of chicken housekeeping genes using next-generation sequencing data.Frontiers in genetics · 2022Article
- N-acetylcysteine Can Induce Massive Oxidative Stress, Resulting in Cell Death with Apoptotic Features in Human Leukemia Cells.International journal of molecular sciences · 2021Article
- Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines.PloS one · 2021Article
- Determination of novel reference genes for improving gene expression data normalization in selected canine reproductive tissues - a multistudy analysis.BMC veterinary research · 2020Article
- Assessment of Enrichment of Human Mesenchymal Stem Cells Based on Plasma and Mitochondrial Membrane Potentials.Bioelectricity · 2020Article
- The role and robustness of the Gini coefficient as an unbiased tool for the selection of Gini genes for normalising expression profiling data.Scientific reports · 2019Article
- Assessment of Aedes albopictus reference genes for quantitative PCR at different stages of development.PloS one · 2018Article
- Dealing with prognostic signature instability: a strategy illustrated for cardiovascular events in patients with end-stage renal disease.BMC medical genomics · 2016Article
- A Novel Non-Immunoglobulin (non-Ig)/BCL6 Translocation in Diffuse Large B-Cell Lymphoma Involving Chromosome 10q11.21 Loci and Review on Clinical Consequences of BCL6 Rearrangements.Pathology oncology research : POR · 2016Review
- Abnormal Expressions of DNA Glycosylase Genes NEIL1, NEIL2, and NEIL3 Are Associated with Somatic Mutation Loads in Human Cancer.Oxidative medicine and cellular longevity · 2016Article
- Article
- The Serum Expression of Selected miRNAs in Pulmonary Sarcoidosis with/without Löfgren's Syndrome.Mediators of inflammation · 2016Article
- Correlation Network Analysis Reveals Relationships between MicroRNAs, Transcription Factor T-bet, and Deregulated Cytokine/Chemokine-Receptor Network in Pulmonary Sarcoidosis.Mediators of inflammation · 2015Article
- Accelerated apoptosis of neutrophils in familial mediterranean Fever.Frontiers in immunology · 2015Article
- Reduced expression of peroxisome proliferator-activated receptor alpha in BAL and blood T cells of non-löfgren's sarcoidosis patients.Journal of inflammation (London, England) · 2015Article
- Normalizing to GADPH jeopardises correct quantification of gene expression in ovarian tumours - IPO8 and RPL4 are reliable reference genes.Journal of ovarian research · 2013Article
- The RHOX homeobox gene cluster is selectively expressed in human oocytes and male germ cells.Human reproduction (Oxford, England) · 2013Article
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Authors and funding
9 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundFor accuracy of quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR), normalisation with suitable reference genes is required. To date, no reference genes have been validated for expression studies of bronchoalveolar (BAL) cells. The aims of this study were to identify gene(s) with stable mRNA expression in BAL cells irrespective of gender, smoking, BAL cellular composition, lung pathology, treatment; and to assess the influence of reference genes on target gene expression data.
resultsThe mRNA expression of ten housekeeping genes (ACTB, ARF1, CANX, G6PD, GAPDH, GPS1, GNB2L1, PSMB2, PSMD2, RPL32) was investigated by qRT-PCR in BAL cells from 71 subjects across a spectrum of lung diseases. The analyses were validated in an independent BAL cohort from 63 sarcoidosis patients and 17 control subjects. A second derivative method was used to calculate expression values (CTt); an equivalence test, applets BestKeeper, geNorm and NormFinder were applied to investigate gene expression stability. Of the investigated genes, PSMB2 (CTt +/- SD, 23.66 +/- 0.86) and RPL32 (18.65 +/- 0.92) were the most stable; both were constantly expressed in BAL samples from parallel investigated cohorts irrespective of evaluated variables. Finally, to demonstrate effect of traditional (ACTB/GAPDH) and novel (PSMB2/RPL32) reference genes as denominators, expression of two cytokines known associated with sarcoidosis was investigated in sarcoid BAL cells. While normalization with PSMB2/RPL32 resulted in elevated IFNG mRNA expression (p = 0.004); no change was observed using GAPDH/ACTB (p > 0.05). CCL2 mRNA up-regulation was observed only when PSMB2/RPL32 were used as denominators (p < 0.03).
conclusionPSMB2 and RPL32 are, therefore, suitable reference genes to normalize qRT-PCR in BAL cells in sarcoidosis, and other interstitial lung disease.
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