A G to T mutation at nucleotide position 2128 in the human FGFR3b coding region resulting in a Cys for Gly substitution (G697C) in the tyrosine kinase domain was observed in 62% (44/ 71) of oral squamous cell carcinomas (OSCC) examined. Immunostained FGFR3b was found in the cytoplasm of prickle cells in normal epithelia, and FGFR3b was localized in the cytoplasm and nucleus in non-FGFR3b mutant OSCC. Overexpressed FGFR3b protein on plasma membranes was noted in OSCC bearing the FGFR3b mutation. Enhanced tyrosine kinase activity of G697C FGFR3b was confirmed. Our results indicate that G697C is an activating mutation causing constitutive ligand-independent FGFR3b signaling. This mutation may be involved in the progression of OSCC and thus the FGFR3b coding sequence may have diagnostic or prognostic value for OSCC. ' 2005 Wiley-Liss, Inc.Key words: FGFR3b; oral squamous cell carcinomas; mutationThe fibroblast growth factor (FGF) family currently includes 22 different gene products that bind to 4 distinct FGF receptors (FGFR1-FGFR4). 1,2 The activities of FGF are mediated by high-affinity FGFR located on the cell surface, which share a common structure of 2 or 3 extracellular immunoglobulinlike loops, a transmembrane domain, and an intracellular split tyrosine kinase domain. 3,4 It is believed that FGFR, when activated by bound FGF in the presence of heparin or heparan sulfate proteoglycan, forms a dimer, which undergoes phosphorylation on tyrosine residues, and transduces signals of cell growth and differentiation. 5,6 It was shown recently that mutations in FGFR cause various congenital skeletal and chondral dysplasia. 7 In these congenital diseases, constitutively active tyrosine kinase activity due to ligand-independent receptor dimerization was observed, and the degree of receptor tyrosine kinase activity correlated with the severity of disease. Squamous cell carcinoma (SCC) is a cancer originating from stratified squamous epithelia that mainly cover skin, oral cavity, esophagus, vagina and bronchus. We have reported previously that all 4 FGFR, including FGFR1 (c isoform), FGFR2 (b isoform), FGFR3 (b isoform) and FGFR4 were expressed exclusively in cells derived from normal oral epithelia and oral squamous cell carcinomas (OSCC). 8 The growth of normal epithelial cells was stimulated by FGF whereas OSCC cell proliferation was not. 9 Mutations in FGFR genes have been reported in human bladder, cervical, gastric and colorectal carcinomas, 10,11 however no such mutations have been reported in OSCC.We examined the mutational status of FGFR3b in a series of human OSCC to determine whether FGFR3b is involved in oral squamous tumorigenesis. We found that 44 OSCC DNAs showed band shifts in exon 17 of the FGFR3b gene by SSCP analysis (Fig. 1a). The DNA sequence of exon 17 in each sample was investigated by direct nucleotide sequencing. A heterozygous missense mutation (G2128T) was observed in exon 17 in all cases (44/71) that exhibited a band shift in PCR-SSCP (Fig. 1b). This mutation resulted in a glycine-tocysteine s...