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Small interfering RNAs suppress the expression of endogenous and GFP-fused epidermal growth factor receptor (erbB1) and induce apoptosis in erbB1-overexpressing cells

MPS-Authors

Nagy,  P.
Max Planck Society;

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Arndt-Jovin,  D. J.
Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society;

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Jovin,  T. M.
Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society;

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Nagy, P., Arndt-Jovin, D. J., & Jovin, T. M. (2003). Small interfering RNAs suppress the expression of endogenous and GFP-fused epidermal growth factor receptor (erbB1) and induce apoptosis in erbB1-overexpressing cells. Experimental Cell Research, 285(1), 39-49. Retrieved from http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6WFC-481FPBR-1-1&_cdi=6791&_user=38661&_pii=S0014482702000502&_orig=search&_coverDate=04%2F15%2F2003&_sk=997149998&view=c&wchp=dGLzVtb-zSkzk&md5=1a576bc18e9242fccd6607d402c15bfe&ie=/sdarticle.pdf.


Cite as: https://hdl.handle.net/11858/00-001M-0000-0012-F11C-4
Abstract
Deregulated and excessive expression of epidermal growth factor receptor (EGFR or erbB1), a transmembrane receptor tyrosine kinase specific for the epidermal growth factor (EGF), is a feature and/or cause of a wide range of human cancers, and thus inhibition of its expression is potentially therapeutic. In RNA interference (RNAi), duplexes of 21-nucleotide RNAs (small interfering RNA, siRNA) corresponding to mRNA sequences of particular genes are used to efficiently inhibit the expression of the target proteins in mammalian cells. Here we show that by using RNAi the expression of endogenous erbB1 can be specifically and extensively (90%) suppressed in A431 human epidermoid carcinoma cells. As a consequence, EGF-induced tyrosine phosphorylation was inhibited and cell proliferation was reduced due to induction of apoptosis. We established an inverse correlation between the level of expressed erbB1 and EGF sensitivity on a cell-by-cell basis using flow cytometry. A431 cells expressing endogenous erbB1 were transfected with erbB1 fused C-terminally to enhanced green fluorescent protein (EGFP). Selective inhibition of the expression of the fusion protein was achieved with an siRNA specific for the EGFP mRNA, whereas the erbB1-specific siRNAs inhibited the expression of both molecules. siRNA-mediated inhibition of erbB I and other erbB tyrosine kinases may constitute a useful therapeutic approach in the treatment of human cancer. (C) 2003 Elsevier Science (USA). All rights reserved.