Reliable detection of p53 aberrations in squamous cell carcinomas of the head and neck requires transcript analysis of the entire coding region

Abstract

Depending on the methodical approach applied, a considerable number of SCCHN may be misclassified regarding their p53 status.

Nuclear survivin expression is associated with HPV-independent carcinogenesis and is an indicator of poor prognosis in oropharyngeal cancer

“…In fact, in the carcinogenesis driven by environmental toxins such as tobacco and alcohol, p53 mutations are more frequently found with subsequent deletion of p53 (Balz et al, 2003). Moreover, biallelic loss of the gene or transcriptional silencing of p53, both of which have been reported in OSCC cell lines, results in a complete loss of transcript in tumour cells (Hauser et al, 2002). These findings might explain the positive correlation of cytoplasmic survivin overexpression and HPV-negative tumours in our series.…”

Comprehensive analysis of the p53 status in mucosal and cutaneous melanomas

“…Primers and PCR conditions are described elsewhere. 28 Analysis of loss of heterozygosity and loss of allele-specific transcription LOH analysis for assessment of allelic imbalance was performed using 3 polymorphic microsatellites localized at or in the vicinity of the p53 locus and the p53 codon 72 polymorphism.…”

Immunological characterization of missense mutations occurring within cytotoxic T cell‐defined p53 epitopes in HLA‐A*0201+ squamous cell carcinomas of the head and neck

“…They did not recognize SaOS-2 cells transfected with either wt p53 or EGFP cDNA. The availability of the HLA-A2 1 UD-SCC 6 SCCHN cell line, which expresses the p53 Y220C mutation, 17,18 facilitated the further characterization of the HLA-A2-restricted p53 mutant p53 Y220C peptide. These effectors had significant reactivity against UD-SSC 6 cells, but none against PCI-13 cells, an HLA-A2 1 SCCHN cell line that expresses a p53 missense mutation at codon 286 and presents the wt p53 217-225 peptide (Fig.…”

“…[17][18][19] T2 and SaOS-2 p53 null cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA). The cell lines were maintained in a complete medium (CM) consisting of RPMI-1640 containing 10% (v/v) FBS, 2 mM L-glutamine, 50 lg/ml streptomycin and 50 IU/ml penicillin (all from Life Technologies, Grand Island, NY).…”

“…The PCR products were purified and sequenced as previously described. 17,18 Transfection of SaOS-2 cells with p53 cDNA Wt p53 and p53 Y220C cDNA were generated from transcripts expressed in normal cells and the UD-SCC 6 cell line, respectively, and ligated into the pAAV-Zeo expression vector under the control of a CMV-promoter, using methods and primers previously described by Balz et al 18 SaOS-2 p53 null cells were transfected with pAAV vectors expressing wt or mutant p53 cDNA, as well as the control vector at a cell density of 0.5 2 3 10 6 /0.1 ml in Nucleofector TM Solution (amaxa biosystems, Cologne, Germany), mixed with 1 5 lg plasmid DNA and transferred into an amaxa certified cuvette. The plasmid DNA transfections were performed by electroporation using a Nucleofector device, according to the manufacture's protocol.…”