MULTIPLE POLYMORPHISMS, BUT NO MUTATIONS, IN THE WAF1 CIP1 GENE IN HUMAN BRAIN-TUMORS/

The cyclin kinase inhibitor WAF1/CIP1, also termed CDKN1, mediates p53 -induced cell cycle arrest in response to DNA damage. This property ma kes it an attractive tumour-suppressor candidate for a p53-associated tumour-suppressor gene. In order to investigate the role of WAF1/CIPI in the pathogenesis of primary human brain tumours we performed single -stranded conformation polymorphism (SSCP) analysis and direct sequenc ing of exon 2 of the gene in a representative series of 158 brain tumo urs and corresponding blood samples. In addition, all tumours were exa mined for mutations in exons 5-8 of the p53 gene. Analysis of WAF1/CIP 1 revealed multiple polymorphisms, the most abundant being AGC-->AGA ( Ser-->Arg) at codon 31 with an allele frequency of 8.5%. Less common p olymorphisms included GTG-->GGG (Val-->Gly) at codon 25, GCC-->ACC (Al a-->Thr) at codon 64, CGC-->CTC (Arg-->Leu) at codon 32, GGC-->AGC (Gl y-->Ser) at codon 14 and GCG-->GTG (Ala-->Val) at codon 39 each with a n allele frequency of 0.3%. These polymorphisms were all located in a conserved region of exon 2. Two of the polymorphisms were also seen in a group of 157 healthy controls indicating that WAF1/CIP1 polymorphis ms do not predispose to cancer. None of the tumours included in our se ries showed a somatic mutation in WAF1/CIP1. All samples were also ana lysed for loss of heterozygosity on the short arm of chromosome 6 in t he region of the WAF1/CIP1 locus. Allelic loss was observed in only on e patient with a glioblastoma. Mutations in the p53 gene were found in 22 of 158 tumours. No association was found between any polymorphism of the WAF1/CIP1 gene, p53 mutations and histopathological tumour type . Our data indicate that WAF1/CIP1 mutations are probably not involved in the formation of primary human brain tumours.