Mutations in the human homologue of the Drosophila segment polarity gene patched in oral squamous cell carcinoma cell lines.
In the present study, we have analyzed tumor deoxyribonucleic acid from oral squamous cell carcinoma (OSCC) cells for patched mutations using an exon-by-exon single strand conformation polymorphism assay and direct sequencing. We found two missense mutations which affected the conserved residue in the transmembrane domains of the gene product and in the intracellular loop at the C-terminal residue implicated in regulating the smoothened molecule. In addition, we demonstrated that the N-terminal fragment of sonic hedgehog (Shh-N) stimulates the growth of normal epithelial cells, the OSCC cell line, NA, and the salivary gland adenocarcinoma cell lines, HSG and HSY, which have no detectable mutation in patched. On the other hand, Shh has no effect on human SCC cells (UE, KA, KO, NI, A431 cells) that have mutations in patched. These results strongly suggest that an Shh-patched signaling is involved in the cell growth of oral epithelial cells and in the tumorigenesis of OSCCs.
Michimukai
E
E
Department of Molecular Oral Medicine & Maxillofacial Surgery 1, Hiroshima University Faculty of Dentistry, Japan.
Kitamura
N
N
Zhang
Y
Y
Wang
H
H
Hiraishi
Y
Y
Sumi
K
K
Hayashido
Y
Y
Toratani
S
S
Okamoto
T
T
D002294:Carcinoma, Squamous Cell / Q000235:genetics*; D002455:Cell Division / Q000187:drug effects; D016895:Culture Media, Serum-Free; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000032:analysis*; D004847:Epithelial Cells / Q000187:drug effects; D053823:Hedgehog Proteins; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D009062:Mouth Neoplasms / Q000235:genetics*; D009154:Mutation; D020125:Mutation, Missense; D000072079:Patched Receptors; D010446:Peptide Fragments / Q000494:pharmacology; D018807:Polymorphism, Single-Stranded Conformational; D012333:RNA, Messenger / Q000032:analysis; D011956:Receptors, Cell Surface; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D015534:Trans-Activators / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D016895:Culture Media, Serum-Free; D004273:DNA, Neoplasm; D053823:Hedgehog Proteins; D008565:Membrane Proteins; D000072079:Patched Receptors; D010446:Peptide Fragments; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; C509239:SHH protein, human; D015534:Trans-Activators