| Project/Area Number |
15590322
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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 |
Human pathology
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| Research Institution | KANAZAWA MEDICAL UNIVERSITY |
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Principal Investigator |
UEDA Yoshimichi Kanazawa Medical University, Dep of Medicine, Professor, 医学部, 教授 (50271375)
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| Co-Investigator(Kenkyū-buntansha) |
FUJITA Takuya Kanazawa Medical University, Dep of Medicine, Lecturer, 医学部, 講師 (40293360)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2004: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | human soft tissue sarcoma / HT1080 / metastasis / cDNA macroarrays / fibronectin 1 gene / junction plakoglobin gene / pulmonary micrometastasis / orthotopic inoculation / ヒト線維肉腫細胞 / 宿主・間質相互応答 / cDNAマイクロアレイ / Plakoglobin遺伝子 / 肺転移 / 悪性線維性組織球腫 / 滑膜肉腫 / c-DNAマクロアレイ / Laser microdissection / 宿主細胞 / 活動性新生血管 |
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| Research Abstract |
1. Selection of genes involved in the tumor-host stroma cell interaction in the metastasis of human fibrosarcoma cell (HT1080)
1) No significant difference of gene expressions between parent and highly metastatic clones of HT1080
Highly metastatic clones were selectively cloned from HT1080 and gene expression profilings were analysed by cDNA macroarrays using Human Cancer 1.2 Atlas Expression Arrays (Clonetech). No significant difference of gene expressions between parent and highly metastatic clones of HT1080 in vitro cultures was found out.
2) Fibronectin 1 (FN 1) gene is overexpressed in HT1080 cells in pulmonary micrometastases
Gene expressions involved in pulmonary micrometastasis-formation were investigated with a lung metastatic model of HT1080 using nude mice injected through tail vein. Several overexpressed and underexpressed genes were demonstrated in pulmonary micrometastasis. Of them, expression of fibronectin 1 (FN 1) gene was increased 50 times compared with that of in vitro-
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cultured HT1080 cells. The prominent upregulation of FN 1 gene transcription was verified by real time RT-PCR and the overexpression was localized in embolized HT1080 tumor cells of the pulmonary micrometastases by laser captured microdissection / real time RT-PCR.
3) Junction plakoglobin gene is down-regulated in orthotopically inoculated intramuscle tumor cells showing enhanced growth and pulmonary micrometastases
Genes involved in the tumor - host stroma cell interaction in the invasion & metastasis of HT1080 cells were investigated using heal pad inoculation (H-group) and intramuscular inoculation (M-group) models in nude mice. M-group showed significantly enhanced tumor cell growth and pulmonary micro-metastases compared with H-group. Gene expressions of tumor cells as well as host stroma cells of M- and H-groups were profiled using both Human Cancer 1.2 Atlas Expression Arrays and Mouse Atlas Expression Arrays. Array-analyses disclosed that expression of junction plakoglobin gene was significantly down-regulated in M-group compared with that of H-group. The down-regulation of junction plakoglobin gene was confirmed by real time RT-PCR at mRNA level and by immunohistochemistry using specific monoclonal antibody to plakoglobin at protein level. No significant genes differently expressed between stroma cells of H- and M-groups were shown.
2. Involvement of the down regulation of junction plakoglobin in human soft tissue sarcomas : Clinical application
Expressions of junction plakoglobin gene were evaluated at both protein and mRNA levels in various kinds of human soft tissue sarcomas.
Junction plakoglobin was overexpressed in synovial sarcoma cells, especially epithelial type. In malignant fibrous histiocytomas (MFH), expressions of junction plakoglobin was significantly down-regulated, at not only protein but also mRNA level, in tumors with pulmonary metastases compared with those without pulmonary metastases. Less
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