| Project/Area Number |
08557085
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
TOGUCHIDA Junya Kyoto University, Research Center for Biomedical Engineering, Associate Professor, 生体医療工学研究センター, 助教授 (40273502)
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| Co-Investigator(Kenkyū-buntansha) |
TABATA Yasuhiko Kyoto University, Research Center for Biomedical Engineering, Associate Professo, 生体医療工学研究センター, 助教授 (50211371)
IKADA Yoshito Kyoto University, Research Center for Biomedical Engineering, Professor, 生体医療工学研究センター, 教授 (00025909)
SASAKI Masao Kyoto University, Radiation Biology Center Professor, 放射線生物研究センター, 教授 (20013857)
NAKAMURA Takashi Kyoto University, Faculty of Medicine, Department of Orthopaedic Surgery, Profes, 医学研究科, 教授 (10201675)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥17,600,000 (Direct Cost: ¥17,600,000)
Fiscal Year 1997: ¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1996: ¥12,400,000 (Direct Cost: ¥12,400,000)
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| Keywords | translocation / liposarcoma / TLS gene / CHOP gene / amisense / gene therapy / 粘液型脂肪肉腫 / 染色体転座 / 融合遺伝子 / アンチセンス療法 |
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| Research Abstract |
1.Genetic analysis of TLS-CHOP translocations in myxoid and round-cell liposarcomas.
Two novel types of fusion transcripts were isolated and sequenced. Precise analysis of genomic fusion points revealed that there are several characteristics on sequences around the breakpoints. Sequence homology with Translin binding site, which has been cloned as a recombination hot spot binding protein in leukemias, was found in two cases. In addition, the breakpoint sequence showed high homology with topoisomerase II binding sites. These results suggested the importance of two proteins in the process of translocation.
2.Introduction of TLS-CHOP fusion genes into preadipocytic cell line.
Coding region of four different types of TLS-CHOP fusion genes were cloned into CMV expression vector and transfected to preadipocytic cell line, Swiss 3T3 L1 cells. Transformed cell lines were established and characterized.
3.Establishment of liposarcoma cell lines expressing TLS-CHOP fusion genes.
Two liposarcoma cell lines were estabilshed. One is KS509, which expressed a novel type of fusion transcript. SV40 transformed derivative (KS509SV) was also established from the parental cell line, and it was used to evaluate the growth inhibitory effects of antisense oligonucleotides (AONs), which were designed to target the fusion point sequences. One of three AONs showed approximately 50% inhibition. The other cell line, KS559, was established from tumors inoculated on SCID mouse. Histological findings of this tumor were compatible with myxoid liposarcomas, and in vitro cells also showed adipocytic morphology, and therefore it was considered to be a suitable model for in vivo and in vitro antisense therapy.
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