| Project/Area Number |
09670208
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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, Department of Medicine, Associate professor, 医学部, 助教授 (50271375)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1998: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1997: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Liposarcoma, well differentiated ; / HMGI-C gene ; / Nested PCR ; / 3'RACE ; / Gene amplification ; / Chromosomal translocation ; / 3'UTR AUUUA motifs / 遺伝子増幅 / ヒト胎児組織 |
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| Research Abstract |
In order to elucidate the molecular mechanism involved in the development and progression of well-differentiated liposarcoma, expression of HMGI-C, molecular mechanism of the activation of HMGI-C gene and functions of HMGI-C in the transformation of fibroblasts were investigated as followings :
1. Expressions of HMGI-C gene at both protein and message levels :
(1) Immunohistochemical and Western blot analyses
(2) Analysis by in situ hybridization (ISH)
2. Molecular mechanism of the activation of HMGI-C gene :
(1) nested reverse transcription polymerase chain reaction (nested RT-PCR)and DNA sequencing
(2) Southern blot analysis for gene amplification
(3) 3' rapid amplifiction of cDNA ends (3'RACE) for chimera gene formation
3. In vitro transformation studies using fibroblasts derived from HMGI-C-/- or HMGI-C +/+ mouse
Tumor cells of well differentiated liposarcoma overexpressed HMGI-C gene (21/21 cases). In contrast, dedifferentiated liposarcomas, which developed in well-differentiated liposarco
… More
ma and are highly malignant, showed loss of HMGI-C expressions. No abnormality was disclosed in the moleular weight of HMGI-C protein expressed in well-differentiated liposarcomas or in cDNA of protein coding region of HMGI-C gene demonstrated by nested RT-PCR.Southern blot analysis did not show any amplification of HMGI-C gene in well differentiated liposarcomas. 3'RACE analysis has not demonstrated any newly formed chimera gene of HMGI-C in well-differentiated liposarcoma so far. In vitro studies showed that fibroblasts derived from HMGI-C -/- mouse transformed more frequently than those derived from HMGI-C -/- mouse, and the transformation in HMGI-C -/- fibroblasts was inhibited by transfection of HMGI-C cDNA.These data clearly demonstrated that activation of HMGI-C gene is involved in the development and progression of well-differentitated liposarcoma. Both in vivo and in vitro data indicated that HMGI-C overexpressed in well-differentiated liposarcomas may inhibit progression to highly malignant tumor, i.e. dedifferentiated liposarcoma. The present data suggested that abnormalies in 3' UTR AUUUA motifs and / or promotor resion might contribute to the activation of HMGI-C in well- differentiated liposarcomas. Less
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