| Project/Area Number |
12470440
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | The University of Tokushima |
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Principal Investigator |
NAGAYAMA Masaru The University of Tokushima, School Of Dentistry, Professor, 歯学部, 教授 (30022867)
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| Co-Investigator(Kenkyū-buntansha) |
MOMOTA Yukihiro The University of Tokushima, School Of Dentistry, Instructor, 歯学部, 助手 (00304543)
FUJISAWA Kenji The University of Tokushima, School Of Dentistry, Instructor, 歯学部, 助手 (40228979)
KAMATA Nobuyuki The University of Tokushima, School Of Dentistry, Associate Professor, 歯学部, 助教授 (70242211)
林 英司 徳島大学, 歯学部・附属病院, 講師 (50173000)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥14,300,000 (Direct Cost: ¥14,300,000)
Fiscal Year 2002: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2001: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2000: ¥8,700,000 (Direct Cost: ¥8,700,000)
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| Keywords | squamous cell carcinoma / protein-free culture / Snail / tumor cell invasion / telomerase / EMT / immortalized human oral keratinocytes / hTERT / 無蛋白培地 / 扁平上皮癌細胞 / 口腔粘膜上皮細胞 / 接着因子 / マトリックスメタロプロテアーゼ / サブトラクション / 増殖制御 / コトリックスメタロプロチアーゼ / 増殖因子 |
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| Research Abstract |
Prognosis of oral squamous cell carcinomas (SOC) depends on tumor cell invasion and metastasis. In order to elucidate the mechanisms of invasion and metastasis, it is important to analyze the functions of cytokines and proteases produced by cancer and the surrounding tissues. Furthermore, it is important to compare characters of SOC cells with normal oral keratinocytes. Cancer cells have been cultured in the media supplemented with fetal bovine serum which are different from the media for normal oral keratinocytes. Also characters of normal cells change by cell divisions with shortening of telomere DNA. Therefore it is necessary to produce protein free medium which support growth of both cancer cells and normal oral keratinocytes and to establish immortalized oral keratinocytes maintaining stable cyto-biological characters as normal cells.
In this study, we first produced a new protein free medium, FLDS. This medium supported the growth of SOC cells and normal oral keratinocytes by cont
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rolling calcium concentration. Furthermore, we demonstrated that SOC cells expressing a transcriptional factor, Snail, which relates to the epithelial-mesenchymal transition (ENT), lacked E-cadherin expression, and were more invasive. Analysis of the effects of EGF and HGF using FLDS medium revealed these two growth factors inhibited only Snail negative and E-cadherin positive SOC cells. Also, we found that expression of matrix metalloproteinase-2 was increased by Snail-induced EMT. Furthermore, after the precise studies of the activity and expressions of telomerase and its associated proteins in normal, premalignant and malignant oral epithelial cells, we succeeded to establish immortalized oral keratinocytes by transfect ion with hTERT, a catalytic component of human telomerase and HPVl6-E7 These cells exhibited normal differentiation abilities in the medium supplemented with EGF and bovine pituitary extract. Our results enable us to compare SOC cells and immortalized oral keratinocytes in the same protein-free medium precisely. Less
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