ArticleJournal of ovarian research2013
Normalizing to GADPH jeopardises correct quantification of gene expression in ovarian tumours - IPO8 and RPL4 are reliable reference genes.
Article in Journal of ovarian research, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 23 citations in OpenAlex.
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- Assessing and validating housekeeping genes in normal, cancerous, and polycystic human ovaries.Journal of assisted reproduction and genetics · 2020Article
- Housekeeping Gene Expression in the Fetal and Neonatal Murine Thymus Following Coxsackievirus B4 Infection.Genes · 2020Article
- Selection and Validation of the Optimal Panel of Reference Genes for RT-qPCR Analysis in the Developing Rat Cartilage.Frontiers in genetics · 2020Article
- Validation of Reference Genes for Gene Expression Normalization in RAW264.7 Cells under Different Conditions.BioMed research international · 2019Article
- Pan-Cancer Analysis of TCGA Data Revealed Promising Reference Genes for qPCR Normalization.Frontiers in genetics · 2019Article
- The microRNA miR-192/215 family is upregulated in mucinous ovarian carcinomas.Scientific reports · 2018Article
- Genomic imbalances are involved in miR-30c and let-7a deregulation in ovarian tumors: implications for HMGA2 expression.Oncotarget · 2017Article
- Consensus reference gene(s) for gene expression studies in human cancers: end of the tunnel visible?Cellular oncology (Dordrecht, Netherlands) · 2015Review
- Reference gene selection for real-time quantitative PCR analysis on ovarian cryopreservation by vitrification in mice.Journal of assisted reproduction and genetics · 2015Article
- Identification of stable reference genes for gene expression analysis of three-dimensional cultivated human bone marrow-derived mesenchymal stromal cells for bone tissue engineering.Tissue engineering. Part C, Methods · 2015Article
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Authors and funding
5 authors at 4 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundTo ensure a correct interpretation of results obtained with quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR), it is critical to normalize to a reference gene with stable mRNA expression in the tissue of interest. GADPH is widely used as a reference gene in ovarian tumour studies, although lacking tissue-specific stability. The aim of this study was to identify alternative suitable reference genes for RT-qPCR studies on benign, borderline, and malignant ovarian tumours.
methodsWe assayed mRNA levels for 13 potential reference genes - ABL1, ACTB, CDKN1A, GADPH, GUSB, HPRT1, HSP90AB, IPO8, PPIA, RPL30, RPL4, RPLPO, and TBP -with RT-qPCR in 42 primary ovarian tumours, using commercially pre-designed RT-qPCR probes. Expression stability was subsequently analysed with four different statistical programs (GeNorm, NormFinder, BestKeeper, and the Equivalence test).
resultsExpression of IPO8, RPL4, TBP, RPLPO, and ACTB had the least variation in expression across the tumour samples according to GeNorm, NormFinder, and BestKeeper. The Equivalence test found variation in expression within a 3-fold expression change between tumour groups for: IPO8, RPL40, RPL30, GUSB, TBP, RPLPO, ACTB, ABL1, and CDKN1A. However, only IPO8 satisfied at a 2-fold change as a cut-off. Overall, IPO8 and RPL4 had the highest, whereas GADPH and HPRT1 the lowest expression stability. Employment of suitable reference genes (IPO8, RPL4) in comparison with unsuitable ones (GADPH, HPRT1), demonstrated divergent influence on the mRNA expression pattern of our target genes - GPER and uPAR.
conclusionsWe found IPO8 and RPL4 to be suitable reference genes for normalization of target gene expression in benign, borderline, and malignant ovarian tumours. Moreover, IPO8 can be recommended as a single reference gene. Neither GADPH nor HPRT1 should be used as reference genes in studies on ovarian tumour tissue.
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