| Project/Area Number |
06454511
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Morphological basic dentistry
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| Research Institution | Okayama University |
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Principal Investigator |
NAGAI Noriyuki Okayama University, School of Dentistry, Professor, 歯学部, 教授 (90085770)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAHARA Kenji Okayama University, School of Dentistry, Associate Professor, 歯学部, 助教授 (50076113)
AKAGI Takumi Okayama University, School of Dentistry, Teaching Assistant, 歯学部, 教務員 (50192878)
MURATA Masaru Okayama University, School of Dentistry, Assistant, 歯学部, 助手 (00260662)
INOUE Masahisa Okayama University, School of Dentistry, Assistant, 歯学部, 助手 (20223274)
NAGATSUKA Hitoshi Okayama University, School of Dentistry, Associate Professor, 歯学部, 助教授 (70237535)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1995: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1994: ¥3,800,000 (Direct Cost: ¥3,800,000)
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| Keywords | Odontogenic tumors / Amelogenin / Ameloblastoma / Adenoid odontogenic tumors / mRNA / Digoxigenin Labeling / Ameloblast / Columnar cells / Ameloblastoma / エナメル上皮腫 / ras蛋白 / c-erbB-2蛋白 / PCNA / p53蛋白 / 悪性エナメル上皮腫 / c-myc蛋白 / 免疫組織化学 |
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| Research Abstract |
The classification of odontogenic tumors is related to the differentiation degree of tumor cells. The localization of amelogenin protein and the expression of amelogenin mRNA are among the indexes of odontogenic tumor cell defferentiation. In our study the probes of amelogenin protein and amelogenin mRNA were made and used to observe their distribution in odontogenic tumors.
Materials and methods : 1.Normal control : Wistar strain rat embryoes at 20 days after gestation were used for investigation of amelogenin protein and mRNA distribution in tooth germ.The specimens were fixed by 4% paraformaldehyde perfusion. 2.Making of probes : Total RNA was extracted from human plexiform type ameloblastoma. Primers (sense : 5-TCGGTCCTGCCTCAT-CACCATCC-3 ; antisense : 5-CCGCTTGGTCTTGTCTGTTG-3) were used for amplification using PCR method. The amplified cDNA was loaded on Vector (Bluescript) and labeled with Dig RNA Labeling Kit (Boehringer Mannheim) to make Digoxigenin labeled single strand RNA prob
… More
e by in vitro transcription. 3.Hybridization : The probe was hybridizad with the specimens at 50^0C for 16 hours. Antigen-antibody reaction was performed using Nucleic acid Detection Kit (Boehringer Mannheim) as to evaluate the hybridization under light microscope. In addition, polyclonal anti-amelogenin antibody was used to localize the distribution of amelogenin protein in odontogenic tumors.
Results and discussion : Both amelogenin protein and mRNA were positive in rat and mouse incisors but negative in undifferentiated enamel epithelia. Amelogenin mRNA was detected frequently in the cells of the central region of the follicles and usually positive in the cytoplasm of columnar cells near to basement membrane while amelogenin protein was occasionally detected only in a few cases of ameloblastomas. These findins suggest that amelogenin mRNA is generally present in ameloblastomas. Amelogenin mRNA was commonly detected in adenoid ondontogenic tumor cells. In addition the localization of amelogenin protein was also observed in adenoid odontogenic tumors. These results proved at molecular level that adenoid odontogenic tumor is a well differentiated tumor. Apart from the above-mentioned investigations immunohistochemistry was also adopted to evaluate cell proliferation, oncogene product and antioncogene product in ameloblastomas. These immunohistochemical studies indicated that immunodetection of PCNA,c-myc protein, ras protein and p53 protein is helpful in evaluating the malignance of ameloblastomas. Less
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