A cell-based high-throughput screen for epidermal growth factor receptor pathway inhibitors.
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.
Lin
Wen-Hsing
WH
Division of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Zhunan, Taiwan.
Song
Jen-Shin
JS
Chang
Teng-Yuan
TY
Chang
Chun-Yu
CY
Fu
Yu-Ning
YN
Yeh
Chi-Ling
CL
Wu
Szu-Huei
SH
Huang
Yu-Wen
YW
Fang
Ming-Yu
MY
Lien
Tzu-Wen
TW
Hsieh
Hsing-Pang
HP
Chao
Yu-Sheng
YS
Huang
Shiu-Feng
SF
Tsai
Shih-Feng
SF
Wang
Lin-Mei
LM
Hsu
John T-A
JT
Chen
Yi-Rong
YR
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004789:Enzyme Activation / Q000187:drug effects; D066246:ErbB Receptors / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D006801:Humans; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects*
D047428:Protein Kinase Inhibitors; D066246:ErbB Receptors