Expression and enzyme activity determination of human cyclooxygenase-1 and-2 in a baculovirus-insect cell system
文献类型:期刊论文
作者 | Zhang, WY; Yang, XN; Jin, DZ; Zhu, XZ |
刊名 | ACTA PHARMACOLOGICA SINICA
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出版日期 | 2004-08 |
卷号 | 25期号:8页码:1000-1006 |
关键词 | baculoviruses prostaglandin-endoperoxide synthase non-steroidal anti inflammatory agents prostaglandins drug inhibition |
ISSN号 | 1671-4083 |
文献子类 | Article |
英文摘要 | AIM: To develop an in vitro intact cell-based assay for screening selective cyclooxygenase inhibitors. METHODS: Human cyclooxygenase-1 (hCOX-1) and cyclooxygenase-2 (hCOX-2) genes were cloned from human monocyte cell line THP-1 cells and expressed in Spodoptera frugiperda (sf9) insect cell line by Bac-to-Bac baculovirus expression systems. Infected sf9 cells were harvested 24 h post-infection (hpi), and distributed to a 24-well plate, preincubated with various nonsteroidal anti-inflammatory drugs, and challenged with 10 mmol/L arachidonic acid; the cyclooxygenase activity was assessed indirectly by prostaglandin E(2)-specific radioimmunoassay. RESULTS: Polymerase chain reaction detection demonstrated that hCOX-1 and hCOX-2 were transposed to the bacmid. Western blot analysis showed that infected sf9 cells could express hCOX-1 and hCOX-2 proteins. Radioimmunoassay demonstrated that both recombinant proteins functioned well in sf9 cells. CONCLUSION: Human cyclooxygenase-1 and cyclooxygenase-2 were successfully expressed in sf9 insect cell line. It can be utilized for the identification of potent and selective inhibitors of hCOX-1 and/or hCOX-2. |
WOS关键词 | SELECTIVE-INHIBITION ; ANTIINFLAMMATORY DRUGS ; SYNTHASE-1 ; BIOSYNTHESIS ; COX-2 |
WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
语种 | 英语 |
CSCD记录号 | CSCD:1845122 |
WOS记录号 | WOS:000223240900005 |
出版者 | SHANGHAI INST MATERIA MEDICA |
源URL | [http://119.78.100.183/handle/2S10ELR8/274038] ![]() |
专题 | 药理学第二研究室 |
通讯作者 | Zhu, XZ |
作者单位 | Chinese Acad Sci, Shanghai Inst Biol Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China |
推荐引用方式 GB/T 7714 | Zhang, WY,Yang, XN,Jin, DZ,et al. Expression and enzyme activity determination of human cyclooxygenase-1 and-2 in a baculovirus-insect cell system[J]. ACTA PHARMACOLOGICA SINICA,2004,25(8):1000-1006. |
APA | Zhang, WY,Yang, XN,Jin, DZ,&Zhu, XZ.(2004).Expression and enzyme activity determination of human cyclooxygenase-1 and-2 in a baculovirus-insect cell system.ACTA PHARMACOLOGICA SINICA,25(8),1000-1006. |
MLA | Zhang, WY,et al."Expression and enzyme activity determination of human cyclooxygenase-1 and-2 in a baculovirus-insect cell system".ACTA PHARMACOLOGICA SINICA 25.8(2004):1000-1006. |
入库方式: OAI收割
来源:上海药物研究所
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