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Effect of Double Transfection of Secretable hSOD and hEGF cDNA on Wound Repair and Cell Damage.

分泌型 hSOD 和 hEGF cDNA 双转染对伤口修复和细胞损伤的影响。

基本信息

批准号：
09672341
负责人：
IWAKAWA Seigo
金额：
$ 1.73万
依托单位：
Kobe Pharmaceutical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672341/
关键词：
Cell Proliferation Epidermal Growth Factor Superoxide Dismutase Gene Transfection Wound Repair Active Oxygen Species

项目摘要

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.

直到去年，我们研究了转染了与信号序列cDNA连接的SOD cDNA的大鼠皮肤成纤维细胞（FR细胞）的超氧化物歧化酶（SOD）的分泌活性。今年，我们比较了幼鼠成纤维细胞（BHK-21细胞）和FR细胞的SOD分泌。通过阳离子脂质体将几种与白介素-2信号序列cDNA连接的SOD cDNA导入BHK-21细胞，使其活性增加。用FR细胞观察，在多次传代过程中，SOD的分泌活性保持不变。而转染不含信号序列cDNA的SOD cDNA的BHK-21细胞培养基中SOD活性低于检测限。这些结果表明，对SOD cDNA进行基因修饰，使其具有分泌SOD的特性，将对哺乳动物的各种细胞有用。现在我们正在研究转染分泌SOD的cDNA的人成纤维细胞分泌SOD的情况。利用BHK-21细胞，研究了细胞内外活性氧对细胞增殖和存活的影响。过氧化氢处理或黄嘌呤/黄嘌呤氧化酶处理改变了活性氧变性酶（SOD、谷胱甘肽过氧化物酶）的活性平衡和细胞活力。低浓度对苯二酚可在细胞内产生活性氧，对细胞增殖的影响不显著。而在低浓度对苯二酚的作用下，成纤维细胞生长因子（FGF）对细胞的增殖作用明显增强，说明细胞内低浓度的活性氧刺激了细胞的增殖。我们正在研究比较在对苯二酚存在或不存在的情况下，引入分泌性表皮生长因子（EGF）的cdna对BHK-21细胞增殖的影响，以研究活性氧对细胞生长的许可作用。