| Project/Area Number |
02670307
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Gastroenterology
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| Research Institution | Okayama University |
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Principal Investigator |
HIGASHI Toshihiro Okayama University, Medical School, Assistant, 医学部付属病院, 助手 (80173136)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1990: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Hepatocellular carcinoma / Melanoma / Metastasis / Stimulative molecule for metastasis / Type IV collagen degrading enzyme / Invasion chamber / ICG / GST-alpha / メラノ-マ / IV型コラ-ゲン分解酵素 / GSTーα |
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| Research Abstract |
I)In vitro model of invasion and metastasis of cultured Hepatoma cells by using the invasion chamber.
The potentiality of hepatoma cells(HuH-7 and PLC/PRF/5) was estimated as follows. We calculated cell numbers in the lower chamber which passed through the 13um pore size chemotaxis filter coated with 50ug of reconstituted matrix gel. The both hepatoma cells did not pass through the chemotaxis filter by themselves. On the contrary, mouse melanoma cells(B16 melanoma cell line) passed through the filter and we recognized that the cells entered into and destroyed the gel. Hepatoma cells could pass through the filter by adding culture medium of melanoma to medium of hepatoma in the upper chamber, indicating that melanoma cells secreted molecules for invasion and metastasis.
II)Analysis of the molecules which stimulate invasion and metastasis of hepatoma cells
Type IV collagen degrading enzyme activity was recognized in the culture medium of both melanoma and PLC/PRF/5. However, this enzyme was found as an inactive form which was activated by APMA. Sephacryl-S 200 gel filtration revealed that the molecular weight of the molecule was 37kD, suggesting that this molecule was different from autocrine motility factor, type IV collagen degrading enzyme and basic FGF.
III)Functional diagnosis of hepatocellular carcinoma
In order to use clinical samples, we established functional diagnostic method for hepatoma. Aspiration biopsy after indocyanine green(ICG) was found as a useful device in which more than 95% of ICG negative tissues were malignant cells. We clarified that the mechanism of ICG loss was partly due to the decrease of cytoplasmic glutathion-S-transferase.
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