Japanese Journal of Clinical Oncology 33:47-50 (2003)
© 2003 Foundation for Promotion of Cancer Research
A New Germline Mutation of the PTCH Gene in a Japanese Patient with Nevoid Basal Cell Carcinoma Syndrome Associated with Meningioma
Department of Surgical Pathology, Showa University Fujigaoka Hospital, Yokohama, Japan
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ABSTRACT |
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 TOP ABSTRACT GENETICS SUMMARYCASE REPORT AND GENETIC...METHODS FOR MUTATION DETECTIONAcknowledgmentsREFERENCES |
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We employed polymerase chain reaction and DNA sequencing analysis to characterize the PTCH gene in a Japanese nevoid basal cell carcinoma syndrome (NBCCS) patient suffering from meningioma, multiple basal cell carcinoma and epidermal cysts. Direct sequence analyses revealed a novel single base deletion at nucleotide 2613 in exon 16 (2613delC) in one PTCH allele, resulting in the frame shift and the introduction of a premature termination codon in this mutated allele.
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GENETICS SUMMARY |
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TOPABSTRACTGENETICS SUMMARYCASE REPORT AND GENETIC...METHODS FOR MUTATION DETECTIONAcknowledgmentsREFERENCES |
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Disorder: Nevoid basal cell carcinoma syndrome (Gorlin syndrome)
Ethnicity: Japanese
Gene: PTCH
GenBank accession number: NT008476, NM000264
Chromosomal assignment: 9q22.3
Type of DNA variant: A germline deletion mutation
Mutation: Deletion of a single nucleotide in exon 16 of the PTCH gene resulting in the frame shift and premature termination codon
Method of mutation detection: PCR/direct sequencing
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CASE REPORT AND GENETIC ANALYSIS |
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TOPABSTRACTGENETICS SUMMARYCASE REPORT AND GENETIC...METHODS FOR MUTATION DETECTIONAcknowledgmentsREFERENCES |
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The nevoid basal cell carcinoma syndrome (NBCCS; MIM 109400), also known as Gorlin syndrome or basal cell nevus syndrome (1–3), is a rare autosomal dominant disorder caused by mutations in the patched gene (PTCH), the human homologue of a Drosophila segment polarity gene, PTCH (4–13). PTCH encodes a transmembrane protein that acts as a negative regulator of hedgehog (HH) signaling. PTCH is a tumor suppressor gene and maps to 9q22.3 in humans. The PTCH protein has 12 hydrophobic membrane-spanning domains, intracellular amino- and carboxy-terminal regions and two large hydrophilic extracellular loops where HH ligand binding occurs.
Since the discovery that PTCH is a gene responsible for NBCCS in 1996, 68 mutations, to our knowledge, have been reported. Mutations of PTCH gene have been found not only in NBCCS patients but also amongst malignant neoplasms such as sporadic basal cell carcinoma (BCC), medulloblastoma, primitive neuroectodermal tumor (PNET), breast cancer, colon cancer and meningioma (14,15). We report here a new single-base deletion in one allele of PTCH gene in a Japanese patient with NBCCS associated with meningioma.
The patient was a 50-year-old Japanese female who had suffered from brain tumor when she was 41 years old. As shown in Fig. 1a, histological diagnosis of this tumor was meningothelial meningioma. Skin excision operations have been carried out 14 times during the past 8 years because of multiple basal cell carcinoma (Fig. 1b). Epidermal cysts have also been resected twice. Clinical features of this patient are summarized in Table 1. Her elder sister has also suffered from skin cancer and her father suffered from adamantinoma and died from skin cancer. Her mother, eldest sister and a younger brother are healthy and have never suffered from skin cancer. Her parents are not consanguineous. The family tree of this patient is shown in Fig. 2. Informed consent for genetic analysis was obtained from the patient and her elder sister.
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We successfully screened this patient with NBCCS for PTCH mutations. As shown in Fig. 1c, mutational analysis of the PTCH gene using a pair of primers, ex16F2 and ex16R2, revealed a heterozygous single nucleotide deletion at nucleotide 2613 in exon 16. A nucleotide sequence of the normal individual is also shown (Fig. 1c). DNA obtained from her elder sister revealed the same mutation at nucleotide 2613 as shown in the patient (data not shown).
Wicking et al. reported that there is no correlation between the position of the premature terminations and the clinical features of NBCCS (10). However, meningioma associated with NBCCS is very rare, ~1% or less (16), and we therefore do not have sufficient data at present to analyze the genotype–phenotype correlation between PTCH mutations and the meningioma associated with NBCCS. Cohen proposed a new hypothesis that with neoplasm arising from NBCCS patients, such as medulloblastoma and meningioma, additional hits are required for carcinogenesis in NBCCS patients (3). Namely, the single germ cell hit results in malformations of NBCCS and the tumor growth occurs by additional hits. In fact, Ling et al. reported that genetic alterations from an additional hit are loss of heterozygosity in the PTCH and p53 loci as well as mutations of p53 gene in NBCCS patients (17). In this case, however, immunohistochemical staining for p53 revealed that meningioma cells were negative for p53 (data not shown).
Table 2 shows PTCH mutations in NBCCS reported so far, which include 63 mutations in exons and five splicing mutations. The present study represents the third mutation of PTCH gene in Japanese NBCCS patients. Analysis of the PTCH gene in NBCCS will provide important information not only for genetic counseling but also for research to examine the correlation between the type of mutations in PTCH gene and clinical features in NBCCS.
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METHODS FOR MUTATION DETECTION |
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TOPABSTRACTGENETICS SUMMARYCASE REPORT AND GENETIC...METHODS FOR MUTATION DETECTIONAcknowledgmentsREFERENCES |
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Genomic DNA was prepared from peripheral blood leukocytes by standard methods using phenol–chloroform extraction and ethanol precipitation. The exons and the intron/exon boundaries in the PTCH gene were analyzed by polymerase chain reaction (PCR) and direct sequencing of the PCR products (18). The primers used for the amplification of the genomic DNA were as follows: PTCH ex16F2, 5'- AGG GTC CTT CTG GCT GCG AG-3'; PTCH ex16R2, 5'- TCA GTG CCC AGC AGC TGG AGT A-3'. Amplified DNA fragments were recovered from a low-melting agarose gel and used for the sequencing analysis.
We sequenced the PCR products directly by using a DNA sequencing system (Model 377; Applied Biosystems). Direct sequencing was performed in both directions and the mutation analysis was repeated independently. Nucleotide numbering is based on GenBank sequence NM000264.
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Acknowledgments |
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TOPABSTRACTGENETICS SUMMARYCASE REPORT AND GENETIC...METHODS FOR MUTATION DETECTIONAcknowledgmentsREFERENCES |
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We thank the patient and her elder sister for making this study possible and Dr Sei for the preparation of the blood samples. This work was supported in part by the High-Technology Center Project of the Ministry of Education, Science, Sports and Culture of Japan (G.T.).
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FOOTNOTES |
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+ For reprints and all correspondence: Genshu Tate, Department of Surgical Pathology, Showa University Fujigaoka Hospital, Fujigaoka 1–30, Aoba-ku, Yokohama 227-8501, Japan
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REFERENCES |
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TOPABSTRACTGENETICS SUMMARYCASE REPORT AND GENETIC...METHODS FOR MUTATION DETECTIONAcknowledgmentsREFERENCES |
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Received August 26, 2002; accepted October 2, 2002
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