Selection of Suitable Reference Genes for Quantitative Real-Time PCR in Apoptosis-Induced MCF-7 Breast Cancer Cells |
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Authors: | Eloise Ferreira Marianne J Cronjé |
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Institution: | (1) Department of Biochemistry, Faculty of Science, University of Johannesburg (APK Campus), P.O. Box 524, Auckland Park, 2006, South Africa; |
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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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Keywords: | |
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