GAPD and tubulin are suitable internal controls for qPCR analysis of oral squamous cell carcinoma cell lines.
The selection of housekeeping genes is critical for gene expression studies. To address this issue, four candidate housekeeping genes, including several commonly used ones, were investigated in oral squamous cell carcinoma cell lines. A simple quantitative RT-PCR approach was employed by comparing relative expression of the four candidate genes within two cancerous cell lines (HN6 and HN31) and one noncancerous cell line (HaCaT) treated or not with EGF and TGF-beta1. Data were analyzed using ANOVA followed by the NormFinder software program. On this basis, stability of the candidate housekeeping genes was ranked and non statistical differences were found using ANOVA test. On the other hand, the NormFinder was able to show that GAPD and TUBB presented the less variable results, representing appropriated housekeeping genes for the samples and conditions analyzed. In conclusion, this study suggests that the GAPD and the TUBB represent adequate normalizers for gene profiling studies in OSCC cell lines, covering, respectively, high and low expression levels genes.
Campos
M S
MS
Oral Pathology Department, School of Dentistry, University of São Paulo, Avenida Prof. Lineu Prestes, 2227 Cidade Universitária, 05508-900 São Paulo, SP, Brazil.
Rodini
C O
CO
Pinto-Júnior
D S
DS
Nunes
F D
FD
D002294:Carcinoma, Squamous Cell / Q000235:genetics*; D045744:Cell Line, Tumor; D020869:Gene Expression Profiling / Q000379:methods*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D006801:Humans; D009062:Mouth Neoplasms / Q000235:genetics*; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D014404:Tubulin / Q000235:genetics*
D014404:Tubulin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases