1996 Fiscal Year Final Research Report Summary
Establishment of A Novel System for Malignancy Grading of Oral Cancer Using Molecular Biological Technique
| Project/Area Number |
05404069
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Surgical dentistry
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| Research Institution | Faculty of Dentistry, Tokyo Medical and Dental University |
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Principal Investigator |
ENOMOTO Shoji Tokyo Medical and Dental University, Faculty of Dentistry, Professor and Chairman, 歯学部, 教授 (40013940)
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| Co-Investigator(Kenkyū-buntansha) |
TACHIKAWA Noriko Tokyo Medical and Dental University, Faculty of Dentistry, Associate professor, 歯学部, 助手 (70236537)
KAMATA Nobuyuki Tokyo Medical and Dental University, Faculty of Dentistry, Assistant professor, 歯学部, 助手 (70242211)
TAKAGI Minoru Tokyo Medical and Dental University, Faculty of Dentistry, Professor and Chairma, 歯学部, 教授 (30014012)
TSUCHIDA Nobuo Tokyo Medical and Dental University, Faculty of Dentistry, Professor and Chairma, 歯学部, 教授 (60089951)
MUROTA Seiitsu Tokyo Medical and Dental University, Faculty of Dentistry, Professor and Chairma, 歯学部, 教授 (50072989)
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| Project Period (FY) |
1993 – 1996
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| Keywords | oral squamous cell carcinoma / normal human keratinocytes from oral mucosa / cell line establishment / vascular endoepitheraial protein / PKC eta / p53 tumor suppressor gene / temperature sensitive mutant / molecular diagnosis |
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| Research Abstract |
To investigate the difference between tumor cells of the primary site and metastasized tumor cells of oral squamous cell carcinoma (OSCC), we tried to establish cell lines using primary and metastasized tumors independently from a patient. In addition, normal human keratinocytes (NHK) from oral mucosa of the same patient was also subjected to cell line establishment, which served as ideal control cells. In 2 out of 5 cell lines, which were successfully established between 1993-1996, the primary and metastasized tumor cell lines and normal kratinocyte cell lines were simultaneously established in the protein free media.
Transcripts from OSCC and NHK cells were differentially displayd by PCR method using sets of arbitrary primers to search for genes specifically expressed in OSCC or NHK cells.
We reported a protein which was expressed by one of the OSCC cell line and was found to suppress the growth of lymphocytes.
In addition, a monoclonal antibody, which recognize an unknown vascular endo
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epitherial protein, were prepared.
PKC eta is a subtype of the PKC is known to be expressed in the epitherial cells. To understand the cell biological behavior, PKC eta was overexpressed in the epithelial cells using an adenovirul vector. It was found that the treatment by the 12-O-tetradecanoylphorbol 13-acetate (TPA) suppressed the growth of epithelial cells, while fibroblasts were not suppressed under the same condition, suggesting a possible gene therapy of OSCC.
We previously reported that p53 tumor suppressor gene is frequently mutated in OSCC,and is considered to be responsible for the development of this tumor. To know the role of the mutation, the p53 mutants found in OSCC were examined by transfection assays. A mutant with Val of codon 138 were found to be a temperature sensitive mutant. To establish a system of molecular diagnosis using the p53 gene mutations, the status of the gene, clinical process and histological malignancy grade are compared in detail. Although the mutation frequency was so high, we could not found an obvious correlation between the existence of mutation and clinical or histological malignancy grade, suggesting that the p53 mutation is important in the initiation of OSCC. Less
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