Please use this identifier to cite or link to this item: https://repositorio.unifesp.br/handle/11600/38884
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dc.contributor.authorLeal, Mariana Ferreira [UNIFESP]
dc.contributor.authorBelangero, Paulo Santoro [UNIFESP]
dc.contributor.authorFigueiredo, Eduardo Antonio [UNIFESP]
dc.contributor.authorCohen, Carina [UNIFESP]
dc.contributor.authorLoyola, Leonor Casilla [UNIFESP]
dc.contributor.authorAndreoli, Carlos Vicente [UNIFESP]
dc.contributor.authorSmith, Marilia Cardoso [UNIFESP]
dc.contributor.authorPochini, Alberto de Castro [UNIFESP]
dc.contributor.authorEjnisman, Benno [UNIFESP]
dc.contributor.authorCohen, Moises [UNIFESP]
dc.date.accessioned2016-01-24T14:40:14Z-
dc.date.available2016-01-24T14:40:14Z-
dc.date.issued2015-03-13
dc.identifierhttp://dx.doi.org/10.1371/journal.pone.0118821
dc.identifier.citationPlos One. San Francisco: Public Library Science, v. 10, n. 3, 16 p., 2015.
dc.identifier.issn1932-6203
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/38884-
dc.description.abstractRotator 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 using 2, 3 or 4 reference genes for most of the analyses. However, COL3A1 expression differed between ruptured and non-ruptured (posterior superior region) tendons of patients only when normalized by HPRT1+TBP+B2M and HPRT1+TBP. On the other hand, the comparison between these two groups using the best trio of reference genes (HPRT1+TBP+ACTB) and 4 reference genes did not revealed a significant difference in COL3A1 expression. Consequently, the use of suitable reference genes for a reliable gene expression evaluation by RT-qPCR should consider the type of tendon samples investigated. HPRT1+TBP+ACTB seems to be the best combination of reference genes for the analysis of involving different tendon samples of individuals with rotator cuff tears.en
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.format.extent16
dc.language.isoeng
dc.publisherPublic Library Science
dc.relation.ispartofPlos One
dc.rightsAcesso aberto
dc.titleIdentification of Suitable Reference Genes for Gene Expression Studies in Tendons from Patients with Rotator Cuff Tearen
dc.typeArtigo
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.description.affiliationUniversidade Federal de São Paulo, Dept Ortopedia & Traumatol, São Paulo, SP, Brazil
dc.description.affiliationUniversidade Federal de São Paulo, Dept Morfol & Genet, Disciplina Genet, São Paulo, SP, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Ortopedia & Traumatol, São Paulo, SP, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Morfol & Genet, Disciplina Genet, São Paulo, SP, Brazil
dc.identifier.fileWOS000351277500046.pdf
dc.identifier.doi10.1371/journal.pone.0118821
dc.description.sourceWeb of Science
dc.identifier.wosWOS:000351277500046
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