| Project/Area Number |
63480309
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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 |
Digestive surgery
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| Research Institution | Kagoshima University |
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Principal Investigator |
SHIMAZU Hisaaki Kagoshima University, 医学部, 教授 (30010242)
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| Co-Investigator(Kenkyū-buntansha) |
村松 寿子 鹿児島大学, 医学部, 助手 (50182134)
愛甲 孝 鹿児島大学, 医学部, 助教授 (60117471)
丸山 征郎 鹿児島大学, 医学部, 助教授 (20082282)
村松 喬 鹿児島大学, 医学部, 教授 (00030891)
高尾 尊身 鹿児島大学, 医学部附属病院, 助手 (80171411)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1990: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1989: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1988: ¥3,500,000 (Direct Cost: ¥3,500,000)
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| Keywords | basement membrane / MATRIGEL / invasion-MTT assay / metastasis / invasion / 血行性転移 / 癌細胞 / 細胞膜 / モノクロ-ナル抗体 / 転移抑制 / Invasion Assay / 再構成基底膜 / MTT assay / レクチン / 細胞膜成分 / モノクローナル抗体 / 内皮細胞 |
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| Research Abstract |
Tumor invasion of basement membrane is a very important step in multi steps of metastasis. Recently, reconstituted basement membrane "MATRIGEL" is used for an in vitro invasion model. Equality and continuity of MATRIGEL coating to the membrane filter was basic condition for the invasion model. Then, we performed the erythrocytes sinking test to examine the condition of MATRIGEL coating. This result indicated that MATRIGEL coating by ordinary methods was unequal and broken. We thought that it was caused by the curved line of the coated MATRIGEL surface. Therefore, we have devised to treat the inner wall of culture plate with paraffin was for the parallel surface of MATRIGEL coating. In addition, we applied colorimetric assay by a tetrazolium saly MTT to the quantitating of invasion cells instead of an ordinary method which was to stain invasion cells and count the number microscopically. As a consequence of these studies, quantitation of the invasion cells was feasible to examine briefly and objectively, and we termed this method invasion-MTT assay. Invasion-MTT assay were performed using various malignant and normal cells. The normal cells revealed almost no invasion, but malignant cells revealed obvious invasion to MATRIGEL. Percent index invasion cells of various malignant cells showed higher value than that of normal cells and its border index was 0.3.
Therefore, it is suggested that invasion-MTT assay is useful and appropriate for the research of tumor metastasis and invasion in relation to the basement membrane.
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