| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1999: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Research Abstract |
In 1998 and 1999, the research project entitled "Regulation of the Expression of Fibrinolysis-related Factors by Inflammatory Cytokines and Oxidative Stress" was performed under the financial support of Grant-in Aid for Scientific Research (C). The results are as follows.
HィイD22ィエD2OィイD22ィエD2 and menadione that can induce reactive oxygen species (ROS) increased the urokinase-type plasminogen activator (uPA) accumulation in paralleled with the induction of uPA mRNA. The half life of uPA mRNA induced by either HィイD22ィエD2OィイD22ィエD2 or menadione was not changed compare to before stimulation, suggesting that ROS would induce uPA by activating the gene transcription. To identify the cis-acting elements in the uPA gene promoter implicated in the ROS-induced uPA gene expression, CAT assay was done using a plasmid containing luciferase reporter gene and 2.4 kbp human uPA promoter. However, by any method using DEAE-dextran, lipofectamin, or electroporation, the transfection efficacy of the plasmid into RC-K8 lymphoma cells was very low and therefore no conclusive results were obtained. Electromobility shifit assay using ィイD132ィエD1P-related consensus oligo DNAs related to the binding sites of AP-1, SP1, NF-kB and CREB, revealed the possible activation of Nf-kB and SP1 in the ROS-stimulated cells. NO is also known as an important chemical mediator affecting the cell function, but any NO donor such as S-nitroso-N-acetyl penicillamine, soium nitroprusside or FK409 influenced the uPA expression. Staphylococcal enterotoxins A and B did not influenced the uPA expression but type 1 and 2 verotoxins induced uPA by activating uPA gene expression. These ROS-induced uPA expressions were also observed in another uPA-producing malignant cells, suggesting that the induction of uPA by ROS would be a general phenomenon observed in a number of cells.
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