Identification of suitable reference genes for gene expression studies in tendons from patients with rotator cuff tear

PloS One
Mariana Ferreira LealMoises Cohen

Abstract

Rotator cuff tear is one of the most common causes of shoulder dysfunction. Gene expression analysis may be a useful tool for understanding tendon tears and the failure of cuff healing, and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) has become an effective method for such studies. However, this technique requires the use of suitable reference genes for data normalization. Here, we evaluate the suitability of six reference genes (18S, ACTB, B2M, GAPDH, HPRT1 and TBP) using samples from the rotator cuff tendons of 28 individuals with tendon tears (3 tendons regions) and 8 controls (2 tendon regions); for the tear patients, we evaluated ruptured and non-ruptured tendon samples. The stability of the candidate reference genes was determined using the NormFinder, geNorm, BestKeeper and DataAssist software packages. Overall, HPRT1 was the best single reference gene, and HPRT1+TBP composed the best pair and HPRT1+TBP+ACTB composed the best trio of reference genes from the analysis of different groups, including the simultaneous analysis of all tissue samples. To identify the optimal combination of reference genes, we evaluated the expression of COL1A1 and COL3A1, and no obvious differences were observed when...Continue Reading

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Citations

Jun 20, 2015·The World Journal of Biological Psychiatry : the Official Journal of the World Federation of Societies of Biological Psychiatry·Marcos Leite SantoroSintia Iole Belangero
Apr 20, 2016·Connective Tissue Research·Paulo Santoro BelangeroMoises Cohen
Apr 4, 2018·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Paulo Santoro BelangeroMariana Ferreira Leal
May 24, 2016·Scientific Reports·Prue M Pereira-FantiniDavid G Tingay
Mar 31, 2020·BMC Musculoskeletal Disorders·Mandeep S VirkBrian J Cole

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Methods Mentioned

BETA
biopsy
electrophoresis
PCR

Software Mentioned

GenEx
Bestkeeper
geNorm
DataAssist
NormFinder

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