Normalization of EGFR mRNA levels following restoration of wild-type p53 in a head and neck squamous cell carcinoma cell line.
Head and neck squamous cell carcinoma (HNSCC) is frequently characterized by mutation of the p53 tumor suppressor gene and increased expression of the epidermal growth factor receptor (EGFR). To determine the potential link between p53 and EGFR expression, we examined the effect of overexpressing wild-type p53 on EGFR mRNA levels in HNSCC. In these studies, a temperature-sensitive (ts) p53 mutant was transfected into an HNSCC line which contained a deletion of one allele of the p53 gene and a mutation of the remaining allele. Following selection, six clones were isolated, characterized by Southern blot analysis, and stable expression of mutant p53 protein was confirmed by immunoblotting of clones grown at the mutant temperature. Total RNA was isolated from transfectants grown at both wild-type or mutant temperatures followed by Northern blot analysis to determine levels of EGFR mRNA expression at the two p53 conformations. Clones grown at the wild-type temperature (32.5 degreesC) demonstrated a 69% 13% decrease in EGFR mRNA compared with the same cells grown at the mutant temperature (39.5 degreesC; p=0.006) indicating that restoration of wild-type p53 reduces EGFR mRNA levels in this HNSCC cell line. These findings suggest that abnormalities in p53 may contribute to activation of EGFR gene transcription.
Department of Otolaryngology, University of Pittsburgh Medical Center, Suite 500, The Eye and Ear Institute Building, Pittsburgh, PA 15213, USA.
J R
JR
Grandis
Q
Q
Zeng
S D
SD
Drenning
D J
DJ
Tweardy
D015152:Blotting, Northern; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D066246:ErbB Receptors / Q000096:biosynthesis*; D016158:Genes, p53; D006258:Head and Neck Neoplasms / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009154:Mutation; D012333:RNA, Messenger / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D015854:Up-Regulation
D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; D066246:ErbB Receptors