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Validation of Suitable Endogenous Control Genes for Quantitative PCR Analysis of microRNA gene expression in a rat model of endometrial cancer
University of Skövde, School of Life Sciences. University of Skövde, The Systems Biology Research Centre.
University of Skövde, School of Life Sciences. University of Skövde, The Systems Biology Research Centre.
University of Skövde, School of Life Sciences. University of Skövde, The Systems Biology Research Centre.
2013 (English)In: Cancer Cell International, ISSN 1475-2867, E-ISSN 1475-2867, Vol. 13, 45Article in journal (Refereed) Published
Abstract [en]

Background

MicroRNAs are small RNA molecules that negatively regulate gene expression by translational inhibition or mRNA cleavage. The discovery that abnormal expression of particular miRNAs contributes to human disease, including cancer, has spurred growing interest in analysing expression profiles of these molecules. Quantitative polymerase chain reaction is frequently used for quantification of miRNA expression due to its sensitivity and specificity. To minimize experimental error in this system an appropriate endogenous control gene must be chosen. An ideal endogenous control gene should be expressed at a constant level across all samples and its expression stability should be unaffected by the experimental procedure.

Results

The expression and validation of candidate control genes (4.5S RNA(H) A, Y1, 4.5S RNA(H) B, snoRNA, U87 and U6) was examined in 21 rat cell lines to establish the most suitable endogenous control for miRNA analysis in a rat model of cancer. The stability of these genes was analysed using geNorm and NormFinder algorithms. U87 and snoRNA were identified as the most stable control genes, while Y1 was least stable.

Conclusion

This study identified the control gene that is most suitable for normalizing the miRNA expression data in rat. That reference gene will be useful when miRNAs expression are analyzed in order to find new miRNA markers for endometrial cancer in rat.

Place, publisher, year, edition, pages
BioMed Central, 2013. Vol. 13, 45
Keyword [en]
Endogenous control genes, microRNA, Endometrial cancer
National Category
Natural Sciences
Research subject
Natural sciences
Identifiers
URN: urn:nbn:se:his:diva-8384DOI: 10.1186/1475-2867-13-45ISI: 000319504800001PubMedID: 23680393Scopus ID: 2-s2.0-84878248686OAI: oai:DiVA.org:his-8384DiVA: diva2:640057
Available from: 2013-08-12 Created: 2013-08-09 Last updated: 2017-12-06Bibliographically approved
In thesis
1. MicroRNA expression profiling in endometrial adenocarcinoma
Open this publication in new window or tab >>MicroRNA expression profiling in endometrial adenocarcinoma
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Place, publisher, year, edition, pages
Örebro: Örebro university, 2015. 53 p.
Series
Örebro Studies in Medicine, ISSN 1652-4063 ; 118
Keyword
Endometrial cancer, microRNA, BDII rat model, normalization, endogenous controls
National Category
Cancer and Oncology
Research subject
Medicine
Identifiers
urn:nbn:se:his:diva-10886 (URN)9789175290638 (ISBN)
Public defence
2015-03-27, Portalen, Högskolan i Skövde, Högskolevägen, 09:00 (English)
Opponent
Supervisors
Available from: 2015-05-04 Created: 2015-05-04 Last updated: 2017-11-27Bibliographically approved

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