@article{APS8328,
author = {Wei-yu ZHANG and Xin-ning YANG and Dao-zhong JIN and Xing-zu ZHU},
title = {Expression and enzyme activity determination of human cyclooxygenase-1 and -2 in a baculovirus-insect cell system},
journal = {Acta Pharmacologica Sinica},
volume = {25},
number = {8},
year = {2016},
keywords = {},
abstract = {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 E2-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.},
issn = {1745-7254}, url = {http://www.chinaphar.com/article/view/8328}
}