Overexpression of the human EGF receptor confers an EGF-dependent transformed phenotype to NIH 3T3 cells

Fiore, Pier Paolo Di; Pierce, Jacalyn H.; Fleming, Timothy P.; Hazan, Rachel B.; Ullrich, Axel; King, C. Richter; Schlessinger, Joseph; Aaronson, Stuart A.

doi:10.1016/0092-8674(87)90592-7

Cited by 605 publications

(372 citation statements)

References 43 publications

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“…Mutations G735S, G796S and E804G increased cell transforming ability of mutated EGFR To assess the transforming potential of these EGFR kinase mutations, anchorage-independent growth assays were performed (Di Fiore et al, 1987;Danielsen and Maihle, 2002). The G735S, G796S and E804G mutants were able to transform NIH3T3 cells to anchorageindependent growth in the absence of exogenous EGF (Figures 3a and b).…”

Section: Construction Of Clonal Nih3t3 Cells Stably Expressing Wild-tmentioning

confidence: 99%

“…Cells stably expressing mutants G735S, E804G and G796S, respectively, showed 12.0-, 11.0-and 5.0-fold increases in their ability to form colonies in suspension media versus cells stably expressing WT EGFR. As previously described (Di Fiore et al, 1987;Greulich et al, 2005), WT EGFR transformed cells after the EGF addition. Upon the addition of EGF to the soft agar medium, there was a significant increase in the number of colonies (Figures 3a and b) of cells expressing WT EGFR and all three mutants G735S, G796S and E804G.…”

Section: Construction Of Clonal Nih3t3 Cells Stably Expressing Wild-tmentioning

confidence: 99%

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Epidermal growth factor receptor activation in prostate cancer by three novel missense mutations

et al. 2008

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While epidermal growth factor receptor (EGFR) dysregulation is known to play a critical role in prostate carcinogenesis, there has been no direct evidence indicating EGFR mutations induce tumorigenesis in prostate cancer. We previously identified four novel EGFR somatic mutations in the EGFR tyrosine kinase domain of prostate cancer patients: G735S, G796S, E804G and R841K. In this study, we investigated the oncogenic potential of these somatic mutations by establishing stable clonal NIH3T3 cells expressing these four mutations and WT EGFR to determine their ability to increase cell proliferation and invasion. In the absence of the EGF ligand, cell proliferation was readily increased in G735S, G796S and E804G mutants compared to WT EGFR. The addition of EGF ligand greatly increased cell growth and transforming ability of these same EGFR mutants. Matrigel invasion assays showed enhanced invasion with G735S, G796S and E804G mutants. Western blot analysis showed that these EGFR mutations enhanced cell growth and invasion via constitutive and hyperactive tyrosine phosphorylation and led to the activation of mitogen-activated protein kinase (MAPK), signal transducer and activator of transcription 3 (STAT3) and Akt pathways. Our findings demonstrate the oncogenic activation of three novel EGFR somatic missense mutations in prostate cancer. Molecules that regulate the mechanisms of their oncogenic activation represent novel targets for limiting tumor cell progression, and further elucidation of these mutations will have utility in prostate cancer treatment.

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Section: Construction Of Clonal Nih3t3 Cells Stably Expressing Wild-tmentioning

confidence: 99%

Section: Construction Of Clonal Nih3t3 Cells Stably Expressing Wild-tmentioning

confidence: 99%

Epidermal growth factor receptor activation in prostate cancer by three novel missense mutations

et al. 2008

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“…Whereas EGFR dimerization is 'autoinhibited' in the absence of EGF, ErbB2 adopts an active-like conformation even without bound ligand (Burgess et al, 2003). Accordingly, simply overexpressing ErbB2 transforms NIH3T3 cells (Hudziak et al, 1987;Di Fiore et al, 1987b), whereas EGFR can only transform NIH3T3 cells in the presence of EGF (Di Fiore et al, 1987a).…”

Section: Introductionmentioning

confidence: 99%

EGF-independent activation of cell-surface EGF receptors harboring mutations found in gefitinib-sensitive lung cancer

2006

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Several somatic mutations within the tyrosine kinase domain of epidermal growth factor receptor (EGFR) have been identified that predict clinical response of non-smallcell lung carcinoma (NSCLC) patients to gefitinib. To test the hypothesis that these mutations cause constitutive EGF receptor signaling, and to investigate its mechanistic basis, we expressed representative examples in a null background and analysed their biochemical properties. Each mutation caused significant EGF-independent tyrosine phosphorylation of EGFR, and allowed the receptor to promote Ba/F3 cell mitogenesis in the absence of EGF, arguing that these are oncogenic mutations. Active mutated receptors are present at the cell surface and are fully competent to bind EGF. Recent structural studies show that the inactive EGFR tyrosine kinase domain is autoinhibited by intramolecular interactions between its activation loop and aC helix. We find that mutations predicted to disrupt this autoinhibitory interaction (including several that have not been described in NSCLC) elevate EGF-independent tyrosine kinase activity, thus providing new insight into how somatic mutations activate EGFR and other ErbB family members.

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“…The EGFR is overexpressed in about 48% of primary human breast tumors (Klijn et al, 1992). Cellular transformation mediated by ligand activation of the EGFR has been experimentally demonstrated in rodent ®broblasts transfected with the EGFR and mammary epithelial cells following the constitutive overproduction of EGF or TGFa (Di Fiore et al, 1987a;Tortora et al, 1989).…”

Section: Introductionmentioning

confidence: 99%

Mammary gland specific hEGF receptor transgene expression induces neoplasia and inhibits differentiation

et al. 2000

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Overexpression of the human EGF receptor confers an EGF-dependent transformed phenotype to NIH 3T3 cells

Cited by 605 publications

References 43 publications

Epidermal growth factor receptor activation in prostate cancer by three novel missense mutations

Epidermal growth factor receptor activation in prostate cancer by three novel missense mutations

EGF-independent activation of cell-surface EGF receptors harboring mutations found in gefitinib-sensitive lung cancer

Mammary gland specific hEGF receptor transgene expression induces neoplasia and inhibits differentiation

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