title: Selection of suitable reference genes for quantitative real-time PCR in apoptosis-induced MCF-7 breast cancer cells.
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.
authors.affiliation: Department of Biochemistry, University of Johannesburg (APK Campus), Auckland Park 2006, South Africa.
authors.forename: Eloise
authors.initials: E
authors.lastname: Ferreira
authors.forename: Marianne J
authors.initials: MJ
authors.lastname: Cronjé
mesh_terms: D048948:14-3-3 Proteins / Q000235:genetics; D000199:Actins / Q000235:genetics; D000704:Analysis of Variance; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D001943:Breast Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology*; D045744:Cell Line, Tumor; D005260:Female; D005819:Genetic Markers; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D050883:HSC70 Heat-Shock Proteins / Q000235:genetics; D018869:Heat-Shock Response / Q000235:genetics; D006657:Histones / Q000235:genetics; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020156:Salicylic Acid / Q000494:pharmacology; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
chemical_list: D048948:14-3-3 Proteins; D000199:Actins; D005819:Genetic Markers; D050883:HSC70 Heat-Shock Proteins; C494371:HSPA8 protein, human; D006657:Histones; C572235:YWHAZ protein, human; D014446:Tyrosine 3-Monooxygenase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D020156:Salicylic Acid