| Project/Area Number |
08671222
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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 |
Hematology
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| Research Institution | TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY |
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Principal Investigator |
NIIYA Kenji TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY,DEPARTMENT CLINICAL LABORATORY MEDICINE,ASSOCIATE PROFESSOR, 医学部, 助教授 (50145116)
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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 |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1996: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | UROKINASE / ANTISENSE / INVASIVENESS / LYMPHOMA CELLS / ELECTROPOLATION / SUBCLONING / uPA / antisense / metastasis / lymphoma cells / invasion / cell culture / matrix / アンチセンス遺伝子 / 遺伝子導入 / 浸潤転移能 / マトリゲル / cAMP |
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| Research Abstract |
Although I have moved to Okayama University Medical School from Toyama Medical and Pharmaceutical University January 1998, the above research is continuously but slowly progressing. A new antisense plasmid covering 540 bp from the translation starting site of uPA gene was constructed by inserting an antisense-oriented cDNA fragment of urokinase gene into a PCI vector, The plasmid was transferred with a plasmid containing a Hygromycin-resistant gene into RC-K8 human pre B lymphoma cells by the electroporation. The cells were cultured in the RPMI- 1640 culture medium containing Hygromycin for 2 months and a number of Hygromycin-resistant clones were established. Among them, a couple of clones exerted a reduced uPA production, approximately a half of that of parent RC-K8 cells. The invasiveness examined using a Matrigel chamber was not significantly different from that of parent cells. These results were unfortunately very similar to that seen in the previous experiments performed in 1996 and 1997. Obviously I could not have the conclusion at present, because I do not suceeed yet in construction of the RC-K8 cells whose uPA production was completely bloc ed. Now I am trying to get the urokinase-nonproducing RC-K8 cells using two different antisense plasmid-vectors and proceeding the research with much more careful election and subcloning.
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