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Selection of Suitable Reference Genes for Quantitative Real-Time PCR in Apoptosis-Induced MCF-7 Breast Cancer Cells

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Abstract

Apoptosis is induced in MCF-7 breast cancer cells following treatment with salicylic acid (20 mM), either in the presence or absence of a heat shock (42°C for 30 min). In order to study the alterations of apoptotic genes with quantitative real-time PCR (qPCR), suitable genes with unchanged expression following the treatments is required for normalizing the gene expression levels. In this study, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-actin (ACTB), Histone H2A (HIST), constitutively expressed heat shock protein 70 (HSC70) and tyrosine 3-monooxygenase/trytophan 5 monooxygenase activation protein, 14-3-3 (YWHAZ) were evaluated as appropriate reference genes. Analysis of gene expression data with one-way ANOVA, geNorm and NormFinder identified HIST and YWHAZ as the least affected during the induction of apoptosis by the different treatments, and is the most suitable gene-pair for normalization during qPCR analysis in MCF-7 breast cancer cells undergoing apoptosis following treatment with SA and/or HS.

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Acknowledgments

This research was supported by the National Research Foundation of South Africa and the Walker Trust Fund of the Faculty of Science, University of Johannesburg, South Africa.

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The authors of this study have no conflicts of interest or any financial disclosures to make.

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  1. Department of Biochemistry, Faculty of Science, University of Johannesburg (APK Campus), P.O. Box 524, Auckland Park, 2006, South Africa

    Eloise Ferreira & Marianne J. Cronjé

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  1. Eloise Ferreira
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  2. Marianne J. Cronjé
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Correspondence to Marianne J. Cronjé.

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Ferreira, E., Cronjé, M.J. Selection of Suitable Reference Genes for Quantitative Real-Time PCR in Apoptosis-Induced MCF-7 Breast Cancer Cells. Mol Biotechnol 50, 121–128 (2012). https://doi.org/10.1007/s12033-011-9425-3

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