1990 Fiscal Year Final Research Report Summary
Study on Epithelial-Mesenchymal Interaction During Promotion Stage of Carcinogenesis
| Project/Area Number |
01570205
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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 |
Experimental pathology
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| Research Institution | Sapporo Medical Collge |
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Principal Investigator |
ENOMOTO Katsuhiko Sapporo Medical Collge, Dept. of Pathology, Associate Professor, 医学部(病理学), 助教授 (20151988)
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| Co-Investigator(Kenkyū-buntansha) |
HATTORI Atsuo Sapporo Medical Collge, Dept. of Pathology, Instructor, 医学部(病理学), 助手 (90208538)
MORI Michio Sapporo Medical Collge, Dept. of Pathology, Professor, 医学部(病理学), 教授 (00045288)
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| Project Period (FY) |
1989 – 1990
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| Keywords | Epithelial-mesenchymal interaction / Carcinogenesis / Co-culture / Gap junction / Tight junction |
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| Research Abstract |
1. Analysis of epithelial-mesenchymal interaction by the co-culture system.
Although the cultured hepatocytes rapidly lost their epithelial morphology, rat hepatocytes co-cultured with bovine aortic endothelial cells maintained their epithelial morphology and trabecular arrangement more than 2 weeks. Rapid appearance of bilecanaliculus like structures and reappearance of intercellular communication ability which was assayed by microinjection of fluorescent dye were also observed in the co-cultured hepatocytes. These suggest that the contact with endothelial cells is important for maintaining the differentiated phenotypes of the cultured hepatocytes.
Co-culture experiments were also carried out using hepatoma cell line HuH7 and hepatoblastoma cell line HuH6, HuH7 did not show any phenotypes of differentiation by the co-culture with endothelial cells, whereas proliferation of endothelial cells was induced. Thus, it is suggested that HuH7 may produce endothelial cell growth factors. On the
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other hand, HuH6 co-cultured with BALB3T3 cells showed reorganization of cell junction structures and reappearance of intercellular communication ability, suggesting the induction of differentiated phenotypes in HuH6 by the co-culture with fibroblasts.
2. Changes of the cell junction molecules during promotion stage of hepatocarcinogenesis.
In order to study alterations of cell junction molecules during rat hepatocarcinogenesis, antibodies against gap junction protein connexin 32 and tight junction protein were generated. Specificity of these antibodies was determined by immunoelectronmicroscopic examination and Western blot analysis. Preneoplastic and neoplastic rat liver lesions induced by an intraperitoneal injection of diethylnitrosamine were examined using these antibodies. The results indicated a great decrease of connexin 32 and tight junction proteins in the hyperplastic nodules and liver cancers. Thus, it is suggested t hat changes of cell junction molecules, which consist of the cell-cell recognition system, occur during promotion stage of hepatocarcinogenesis. Less
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