Effect of Double Transfection of Secretable hSOD and hEGF cDNA on Wound Repair and Cell Damage.
Project/Area Number |
09672341
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
応用薬理学・医療系薬学
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Research Institution | Kobe Pharmaceutical University |
Principal Investigator |
IWAKAWA Seigo Kobe Pharmaceutical University, Depertment of Pharmacy, Professor, 薬学部, 教授 (50168548)
|
Co-Investigator(Kenkyū-buntansha) |
HIRAI Midori Kobe Pharmaceutical University, Depaertment of Pharmacy, Associate professor, 薬学部, 助教授 (70228766)
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Project Period (FY) |
1997 – 1998
|
Project Status |
Completed (Fiscal Year 1998)
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Budget Amount *help |
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥1,800,000 (Direct Cost: ¥1,800,000)
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Keywords | Cell Proliferation / Epidermal Growth Factor / Superoxide Dismutase / Gene Transfection / Wound Repair / Active Oxygen Species |
Research Abstract |
Until last year we investigated the secretory activity of superoxide dismutase (SOD) in rat skin fibroblasts (FR cells) transfected with cDNA of SOD connecting with a signal sequence cDNA.This year we compared the secretion of SOD from Baby Hamster fibroblasts (BHK-21 cells) with that from FR cells. Introduction of several cDNAs of SOD connecting with a signal sequence cDNA of interleukin-2 by cationic liposomes to BHK-21 cells increased the activity of SOD in the incubation medium. As observed by the use of FR cells, the secretory activity of SOD was maintained during the several passages. However, the SOD activity in the medium from BHK-21 cells transfected with cDNA of SOD without the signal sequence cDNA was below the detection limit. These results indicated this genetic modification of SOD cDNA to give secretory property for SOD would be useful for various cells from mammalian species. Now we are studying the secretion of SOD from human fibroblasts trasfected with the cDNA of the secretory SOD. The effect of active oxygens outside and/or inside cells on cell proliferation and survival was investigated by the use of BHK-21 cells. The treatment with hydrogen peroxide or xanthine/xanthine oxidase changed the activity balance of active oxygen degenerating enzymes, such as SOD and glutathione peroxidase, and cell viability. Low concentration of hydroquinone, which produces active oxygen inside the cells, showed insignificant effect on cell proliferation. However, the cell proliferation by fibroblast growth factor (FGF) was significantly increased in the presence of the low concentration of hydroquinone, indicating the low concentration of active oxygen inside cells stimulates the cell proliferation. We are investigating to compare the effect of introduction of cDNAof secretory epidermal growth factor (EGF) to the BHK-21 cells on cell proliferation in the presence or absence of hydroquinone to study a permissive effect of active oxygen for cell growth.
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Report
(3 results)
Research Products
(7 results)