Construction of a Transcription Factor Based Drug Screening System and the Molecular Mechanism of PGE1-mediated NF-IL6 Activation

• Main Authors: Po-wen Huang, 黃柏文
• Other Authors: Chiun-Jye Yuan
• Format: Others
• Language: en_US
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/36788651591460223704

碩士 === 國立交通大學 === 生物科技研究所 === 91 === The screening for drugs in regulating the host immune and inflammatory response is important for the therapeutical purpose. Among the known transcription factors, NF-IL6 and NF-kappaBplay pivotal roles in various physiological events such as the inflammatory response, immunity, differentiation, proliferation, tumorgenesis, and numerous other cellular responses. Hence, NF-IL6- and NF-kappaB-based drug screening systems were constructed in our laboratory for the screening of lead components from Chinese herbs for the regulation of immune and inflammatory responses. The p3X-NIL-Lucneo or p4X-NkB-Lucneo stably transfected RAW264.7 cell clones were generated and used for screening of corresponding agonists from various traditional Chinese herbs. We found that the n-butanol layer of Gu Jing Cao could significantly induce the luciferase activity of RAWNFkB cells as well as RAWNFIL6 cells compared with that of untreated cells. In previous studies, we have demonstrated that PGE1 could mediate NF-IL6 activation in HEK293 cells. In this study, we further demonstrate that PGE1-mediated NF-IL6 activation in HEK293 might be controlled by a PKA-dependent manner. The induction of luciferase activity in HEK293 cells was not dependent on the NF-IL6 gene expression or nuclear translocation. Interestingly, the IL-6 mRNA was increased after treatment of PGE1 possibly via the activation of NF-IL6.