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Defining suitable reference genes for RT-qPCR analysis on human sertoli cells after 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure

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Abstract

Quantitative real-time RT-PCR (qPCR) has proven to be a valuable molecular technique to quantify gene expression. There are few studies in the literature that describe suitable reference genes to normalize gene expression data. Studies of transcriptionally disruptive toxins, like tetrachlorodibenzo-p-dioxin (TCDD), require careful consideration of reference genes. The present study was designed to validate potential reference genes in human Sertoli cells after exposure to TCDD. 32 candidate reference genes were analyzed to determine their applicability. geNorm and NormFinder softwares were used to obtain an estimation of the expression stability of the 32 genes and to identify the most suitable genes for qPCR data normalization.

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Acknowledgments

Mariana Antunes Ribeiro was a FAPESP fellowship’s recipient (2011/04202-1) and Wellerson Rodrigo Scarano was recipient of support research from FAPESP (2012/00253-3).

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Department of Morphology, Institute of Biosciences, UNESP- Univ Estadual Paulista, Botucatu, SP, 18618-970, Brazil

    Mariana Antunes Ribeiro, Leonardo Nazário de Moraes & Wellerson Rodrigo Scarano

  2. Department of Genetics, Institute of Biosciences, UNESP- Univ Estadual Paulista, Botucatu, SP, 18618-970, Brazil

    Mariana Bisarro dos Reis & Cláudia Aparecida Rainho

  3. Utah Fertility Center, Pleasant Grove, UT, 84062, USA

    Christine Briton-Jones

Authors
  1. Mariana Antunes Ribeiro
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  2. Mariana Bisarro dos Reis
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  3. Leonardo Nazário de Moraes
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  4. Christine Briton-Jones
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  5. Cláudia Aparecida Rainho
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  6. Wellerson Rodrigo Scarano
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Correspondence to Wellerson Rodrigo Scarano.

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Ribeiro, M.A., dos Reis, M.B., de Moraes, L.N. et al. Defining suitable reference genes for RT-qPCR analysis on human sertoli cells after 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure. Mol Biol Rep 41, 7063–7066 (2014). https://doi.org/10.1007/s11033-014-3637-0

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  • DOI: https://doi.org/10.1007/s11033-014-3637-0

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