| Project/Area Number |
17591422
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Digestive surgery
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| Research Institution | Sapporo Medical University |
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Principal Investigator |
YAMAGUCHI Koji Sapporo Medical University, School of Medicine, Assistant Professor, 医学部, 講師 (60315512)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA Yasutoshi Sapporo Medical University, School of Medicine, Assistant, 医学部, 講師 (80311893)
HIRATA Koichi Sapporo Medical University, School of Medicine, Professor, 医学部, 教授 (50136959)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | gastric cancer / orthotopic implantation / VEGF-C gene / lymph node metastasis / cDNA microarray |
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| Research Abstract |
To investigate the molecular mechanisms by which carcinoma cells metastasize, we have established liver metastatic models. Human gastric carcinoma cell line AZ521 (5 × 10^6/0.1ml) was transplanted orthotopically into the stomach of nude mice using cell suspension technique. Athymic female BALB/c nu/nu mice, 6-7 weeks old, weighing 18-22g were used. After from 6 to 8 weeks, the liver and regional lymph nodes with a few metastatic foci of AZ521 cells were dissected, we established AZH1G and AZL5G. The same procedure was repeated using AZH1G and AZL1G, and AZH5G and AZL5G was established upon the fifth cycle of stepwise selection. We have also established AZ3P3G using intraperitoneally and orthotopically stepwise selection methods. 1) AZL5G produced lymph node mestastasis in 85.0%, and MKNL3G produced lymph node mestastasis in 60.0% of mice. 2) Each metastatic cell grew significantly larger than parental AZ521 in vivo. 3) A cell motility assay demonstrated that the migration ability of each metastatic cell was clearly higher than that of parental cell lines. 4) The expression of several cell adhesion molecules by FACS analysis showed that the incidence of α_1 and α_2 integrins expression in AZL5G cells was significantly higher than in AZ521. Then, MKNL3G cells expressed suggestive up-regulation of α_2 integrin. Moreover, Comprehensive analysis of the biological behavior of highly metastatic cell lines and the less metastatic same parental lines were demonstrated the differences in adhesive activity and VEGF production. Up-regulated genes were endothelin-A receptor and TGF-beta II R alpha in AZL5G. Above all, our experimental models should be useful for investigation of the mechanisms of metastasis in human gastric carcinoma. Up-regulated genes in MKNL3G and inhibitory methods are currently underway.
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