Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10532911 | Analytical Biochemistry | 2008 | 6 Pages |
Abstract
Epidermal growth factor receptor (EGFR) is a valid drug target for development of target-based therapeutics against non-small-cell lung cancer. In this study, we established a high-throughput cell-based assay to screen for compounds that may inhibit EGFR activation and/or EGFR-mediated downstream signaling pathway. This drug screening platform is based on the characterization of an EGFR-transfected 32D cell line (32D-EGFR). The expression of EGFR in 32D cells allowed cell proliferation in the presence of either epidermal growth factor (EGF) or interleukin 3 (IL-3) and provided a system for both screening and counterscreening of EGFR pathway-inhibitory compounds. After the completion of primary and secondary screenings in which 32D-EGFR cells were grown under the stimulation of either EGF or IL-3, 9 of 20,000 compounds were found to selectively inhibit the EGF-dependent proliferation, but not the IL-3-dependent proliferation, of 32D-EGFR cells. Subsequent analysis showed that 3 compounds of the 9 initial hits directly inhibited the kinase activity of recombinant EGFR in vitro and the phosphorylation of EGFR in H1299 cells transfected with EGFR. Thus, this 32D-EGFR assay system provides a promising approach for identifying novel EGFR and EGFR signaling pathway inhibitors with potential antitumor activity.
Keywords
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Wen-Hsing Lin, Jen-Shin Song, Teng-Yuan Chang, Chun-Yu Chang, Yu-Ning Fu, Chi-Ling Yeh, Szu-Huei Wu, Yu-Wen Huang, Ming-Yu Fang, Tzu-Wen Lien, Hsing-Pang Hsieh, Yu-Sheng Chao, Shiu-Feng Huang, Shih-Feng Tsai, Lin-Mei Wang, John T.-A. Hsu, Yi-Rong Chen,