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Optimal reference genes for normalization of qPCR gene expression data from proton and photon irradiated dermal fibroblasts
Stockholm University, Faculty of Science, Department of Physics.ORCID iD: 0000-0002-4160-1078
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Number of Authors: 82018 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 8, article id 12688Article in journal (Refereed) Published
Abstract [en]

The transcriptional response of cells exposed to proton radiation is not equivalent to the response induced by traditional photon beams. Changes in cellular signalling is most commonly studied using the method Quantitative polymerase chain reaction (qPCR). Stable reference genes must be used to accurately quantify target transcript expression. The study aim was to identify suitable reference genes for normalisation of gene expression levels in normal dermal fibroblasts irradiated with either proton or photon beams. The online tool RefFinder was used to analyse and identify the most stably expressed genes from a panel of 22 gene candidates. To assess the reliability of the identified reference genes, a selection of the most and least stable reference genes was used to normalise target transcripts of interest. Fold change levels varied considerably depending on the used reference gene. The top ranked genes IPO8, PUM1, MRPL19 and PSMC4 produced highly similar target gene expression, while expression using the worst ranked genes, TFRC and HPRT1, was clearly modified due to reference gene instability.

Place, publisher, year, edition, pages
2018. Vol. 8, article id 12688
National Category
Radiology, Nuclear Medicine and Medical Imaging Physical Sciences
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URN: urn:nbn:se:su:diva-160081DOI: 10.1038/s41598-018-30946-0ISI: 000442530000051PubMedID: 30139945OAI: oai:DiVA.org:su-160081DiVA, id: diva2:1251819
Available from: 2018-09-28 Created: 2018-09-28 Last updated: 2019-12-09Bibliographically approved

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