A cell-based high-throughput screen for epidermal growth factor receptor pathway inhibitors |
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Authors: | Lin Wen-Hsing Song Jen-Shin Chang Teng-Yuan Chang Chun-Yu Fu Yu-Ning Yeh Chi-Ling Wu Szu-Huei Huang Yu-Wen Fang Ming-Yu Lien Tzu-Wen Hsieh Hsing-Pang Chao Yu-Sheng Huang Shiu-Feng Tsai Shih-Feng Wang Lin-Mei Hsu John T-A Chen Yi-Rong |
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Institution: | a Division of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Zhunan, Taiwan b Division of Molecular & Genomic Medicine, National Health Research Institutes, Zhunan, Taiwan c Department of Chemical Engineering, National Tsing-Hua University, Hsin-Chu, Taiwan |
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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. |
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Keywords: | EGFR 32D cell line HTS NSCLC TKI |
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