Analysis of alterations of WFDC1, a new putative tumour suppressor gene, in hepatocellular carcinoma.
WFDC1 is a recently isolated human gene identified as a tumour suppressor gene candidate. It is not known whether alterations in this gene are associated with human cancers. The WFDC1 gene maps in human chromosome 16q24, an area of frequent loss of heterozygosity (LOH) in several tumour types, in particular in hepatocellular carcinoma (HCC). We investigated its role in 46 European HCC by means of the detection of LOH at the WFDC1 locus. We describe here an assay for the detection of loss of heterozygosity at this locus using two dinucleotide repeat polymorphisms identified in WFDC1 introns, with a combined informativity of 86%. LOH was observed in 4/40 informative HCC samples. We further investigated the role of WFDC1 as a tumour suppressor gene candidate in five hepatocellular cell lines and in tumours exhibiting LOH by means of mutation, promoter methylation and gene expression analysis. In HCC samples showing LOH, no mutation of the remaining allele was observed. No significant up or down gene expression was observed in tumour samples comparatively to normal liver and gene expression did not seem related to promoter methylation. These results suggest a minor role, if any, of WFDC1 in hepatocarcinogenesis. However, this approach might be useful for investigating the role of this candidate tumour suppressor gene in other tumour types.
Saffroy
Raphaël
R
Service de Biochimie et Biologie moléculaire, Hôpital Universitaire Paul Brousse, UPRES 1596-Faculté de Médecine Paris-Sud, 14 avenue Paul Vaillant Couturier 94804 Villejuif Cedex, France. raphael.saffroy@pbr.ap-hop-paris.fr
Riou
Philippe
P
Soler
Gwendoline
G
Azoulay
Daniel
D
Emile
Jean-François
JF
Debuire
Brigitte
B
Lemoine
Antoinette
A
D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D002478:Cells, Cultured; D002885:Chromosomes, Human, Pair 16 / Q000235:genetics; D019175:DNA Methylation; D004252:DNA Mutational Analysis; D018900:Dinucleotide Repeats / Q000235:genetics; D016147:Genes, Tumor Suppressor; D022781:Hepatocytes; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics*; D019656:Loss of Heterozygosity / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D011506:Proteins; C484310:WFDC1 protein, human