ArticleBMC molecular biology2011
Selection of reference genes for gene expression studies in ultraviolet B-irradiated human skin fibroblasts using quantitative real-time PCR.
Article in BMC molecular biology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 24 citations in OpenAlex.
- A Strategy for the Selection of RT-qPCR Reference Genes Based on Publicly Available Transcriptomic Datasets.Biomedicines · 2023Article
- Nanoencapsulation of Pomegranate Extract to Increase Stability and Potential Dermatological Protection.Pharmaceutics · 2021Article
- Exploring the effect of UV-C radiation on earthworm and understanding its genomic integrity in the context of H2AX expression.Scientific reports · 2020Article
- Article
- Selecting suitable reference genes for qPCR normalization: a comprehensive analysis in MCF-7 breast cancer cell line.BMC molecular and cell biology · 2020Article
- MET ectodomain shedding is associated with poor disease-free survival of patients diagnosed with oral squamous cell carcinoma.Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc · 2020Article
- An Integrated Preprocessing Approach for Exploring Single-Cell Gene Expression in Rare Cells.Scientific reports · 2019Article
- Misleading Westerns: Common Quantification Mistakes in Western Blot Densitometry and Proposed Corrective Measures.BioMed research international · 2019Article
- Transcriptome-wide identification of optimal reference genes for expression analysis of Pyropia yezoensis responses to abiotic stress.BMC genomics · 2018Article
- Evaluation of endogenous control gene(s) for gene expression studies in human blood exposed to 60Co γ-rays ex vivo.Journal of radiation research · 2015Article
- Immunological visibility: posttranscriptional regulation of human NKG2D ligands by the EGF receptor pathway.Science translational medicine · 2014Article
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Authors and funding
5 authors at 2 institutions in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
backgroundReference genes are frequently used to normalise mRNA levels between different samples. The expression level of these genes, however, may vary between tissues or cells and may change under certain circumstances. Cytoskeleton genes have served as multifunctional tools for experimental studies as reference genes. Our previous studies have demonstrated that the expression of vimentin, one cytoskeletal protein, was increased in ultraviolet B (UVB)-irradiated fibroblasts. Thus, we examined the expression of other cytoskeleton protein genes, ACTB (actin, beta), TUBA1A (tubulin, alpha 1a), and TUBB1 (tubulin, beta 1), in human dermal fibroblasts irradiated by UVB to determine which of these candidates were the most appropriate reference genes.
resultsQuantitative real-time PCR followed by analysis with the NormFinder and geNorm software programmes was performed. The initial screening of the expression patterns demonstrated that the expression of VIM was suppressed after UVB irradiation at doses ≥25 mJ/cm(2) and that the expression of TUBA1A was significantly reduced by UVB doses ≥75 mJ/cm(2) in cultured human dermal fibroblasts. The analysis of the experimental data revealed ACTB to be the most stably expressed gene, followed by GAPDH (aglyceraldehyde-3-phosphate dehydrogenase), under these experimental conditions. By contrast, VIM was found to be the least stable gene. The combination of ACTB and TUBB1 was revealed to be the gene pair that introduced the least systematic error into the data normalisation.
conclusionThe data herein provide evidence that ACTB and TUBB1 are suitable reference genes in human skin fibroblasts irradiated by UVB, whereas VIM and TUBA1A are not and should therefore be excluded as reference genes in any gene expression studies involving UVB-irradiated human skin fibroblasts.
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