Identification of Suitable Reference Genes for Gene Expression Studies in Tendons from XXXX with Rotator Cuff Tear.

@entity1 @entity1414 @entity7501 @entity924 @entity409 @entity2262 @entity13239 @entity11567 @entity14875 @entity9996
UNASSIGNED: Rotator cuff tear is one of the most common causes of @entity409 . Gene expression analysis may be a useful tool for understanding @entity924 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, @entity14875 , @entity2262 , @entity7501 , @entity9996 and @entity11567 ) using samples from the rotator cuff tendons of 28 individuals with @entity924 (3 tendons regions) and 8 controls (2 tendon regions); for the tear @entity1 , 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, @entity9996 was the best single reference gene, and @entity9996 + @entity11567 composed the best pair and @entity9996 + @entity11567 + @entity14875 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 @entity13239 and @entity1414 , and no obvious differences were observed when using 2, 3 or 4 reference genes for most of the analyses. However, @entity1414 expression differed between ruptured and non-ruptured (posterior superior region) tendons of @entity1 only when normalized by @entity9996 + @entity11567 + @entity2262 and @entity9996 + @entity11567 . On the other hand, the comparison between these two groups using the best trio of reference genes ( @entity9996 + @entity11567 + @entity14875 ) and 4 reference genes did not revealed a significant difference in @entity1414 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. @entity9996 + @entity11567 + @entity14875 seems to be the best combination of reference genes for the analysis of involving different tendon samples of individuals with rotator cuff tears.
