title: TP53 mutational pattern in Spanish and Polish non-small cell lung cancer patients: null mutations are associated with poor prognosis.
abstract: Inactivation of TP53 tumor suppressor gene is the most frequent molecular alteration in NSCLC, involving up to 60% of cases. Furthermore, TP53 mutational spectrum is related to the type of mutagen exposure, as well as racial and/or diet differences. Nearly 95% of TP53 perturbations affect codons included within exons 5-8 which encode for almost the entire DNA-binding domain. In this study we addressed the possible prognostic value of the molecular alterations identified in exons 5-8 of the TP53 gene in DNAs from 151 paraffin-embedded NSCLC sections corresponding to 59 Spanish and 92 Polish stage I-IIIA resected patients. PCR/single-strand conformation polymorphism (SSCP) analysis revealed that the occurrence of TP53 exon 5-8 mutations was 17/59 (29%) in the Spanish cohort and 17/92 (18%) in the Polish group. However, when DNA sequencing analysis was performed, these frequencies were reduced because of the presence of SSCP-false positive, intronic and silent mutations and polymorphisms. Fifteen of the 59 Spanish NSCLC tumors (25%) harbored TP53 mutations affecting exons 5-8 coding sequences, whereas only 12 of 92 Polish neoplasms (13%) contained alterations in the central hydrophobic region of p53. Our results indicate that the occurrence of TP53 mutations affecting exon 5-8 coding sequences in some European NSCLC populations may be lower than previously reported, and that the TP53 mutational patterns of these cohorts differ somewhat. The Spanish NSCLC patients contained missense mutations (9/59, 15%) and a relatively high percentage of null mutations (5/59, 8%) while the Polish patients mostly harbored missense mutations (9/92, 10%) and only one tumor contained a null type (1/92, 1%). Moreover, most TP53 missense mutations in the Spanish group were located outside the conserved regions, whereas the same mutations in the Polish group affected conserved amino acids. Furthermore, the Polish patients harbored a high percentage of G-->A transitions (most of them at non-CpG sites), while G-->T transversions were predominant in the Spanish group. Our findings suggest that there may be different racial or exogenous factors in these two populations which may help to explain both the distinct TP53 mutational pattern and the lower frequency obtained in the Polish group. The presence of missense mutations did not confer a worse clinical outcome in these subsets of NSCLC patients. However, patients whose tumors contained null TP53 gene mutations had a 5 month median disease-free survival time in contrast with 42 months in those patients without mutations (P=0.008). These findings suggest that loss of p53 function may enhance tumor progression in NSCLC patients independently of whether dominant negative TP53 missense mutations are present.
authors.affiliation: Laboratory of Molecular Biology of Cancer, Universitat Autònoma de Barcelona, Hospital Universitari Germans Trias i Pujol, Catalonia, Spain.
authors.forename: J M
authors.initials: JM
authors.lastname: de Anta
authors.forename: E
authors.initials: E
authors.lastname: Jassem
authors.forename: R
authors.initials: R
authors.lastname: Rosell
authors.forename: M
authors.initials: M
authors.lastname: Martínez-Roca
authors.forename: J
authors.initials: J
authors.lastname: Jassem
authors.forename: E
authors.initials: E
authors.lastname: Martínez-López
authors.forename: M
authors.initials: M
authors.lastname: Monzó
authors.forename: J J
authors.initials: JJ
authors.lastname: Sánchez-Hernández
authors.forename: I
authors.initials: I
authors.lastname: Moreno
authors.forename: M
authors.initials: M
authors.lastname: Sánchez-Céspedes
mesh_terms: D000368:Aged; D019943:Amino Acid Substitution / Q000235:genetics; D002289:Carcinoma, Non-Small-Cell Lung / Q000453:epidemiology / Q000235:genetics*; D004252:DNA Mutational Analysis; D018572:Disease-Free Survival; D005260:Female; D016368:Frameshift Mutation; D017353:Gene Deletion; D005787:Gene Frequency; D016158:Genes, p53; D006801:Humans; D008175:Lung Neoplasms / Q000453:epidemiology / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D011044:Poland / Q000453:epidemiology; D011379:Prognosis; D013030:Spain / Q000453:epidemiology