RET/PTC-induced cell growth is mediated in part by epidermal growth factor receptor (EGFR) activation: evidence for molecular and functional interactions between RET and EGFR.
RET/PTC rearrangements are one of the genetic hallmarks of papillary thyroid carcinomas. RET/PTC oncoproteins lack extracellular or transmembrane domains, and activation takes place through constitutive dimerization mediated through coiled-coil motifs in the NH(2) terminus of the chimeric protein. Based on the observation that the epidermal growth factor receptor (EGFR) kinase inhibitor PKI166 decreased RET/PTC kinase autophosphorylation and activation of downstream effectors in thyroid cells, despite lacking activity on the purified RET kinase, we proceeded to examine possible functional interactions between RET/PTC and EGFR. Conditional activation of RET/PTC oncoproteins in thyroid PCCL3 cells markedly induced expression and phosphorylation of EGFR, which was mediated in part through mitogen-activated protein kinase signaling. RET and EGFR were found to coimmunoprecipitate. The ability of RET to form a complex with EGFR was not dependent on recruitment of Shc or on their respective kinase activities. Ligand-induced activation of EGFR resulted in phosphorylation of a kinase-dead RET, an effect that was entirely blocked by PKI166. These effects were biologically relevant, as the EGFR kinase inhibitors PKI166, gefitinib, and AEE788 inhibited cell growth induced by various constitutively active mutants of RET in thyroid cancer cells as well as NIH3T3 cells. These data indicate that EGFR contributes to RET kinase activation, signaling, and growth stimulation and may therefore be an attractive therapeutic target in RET-induced neoplasms.
Division of Endocrinology and Metabolism, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA.
Michelle
M
Croyle
Nagako
N
Akeno
Jeffrey A
JA
Knauf
Doriano
D
Fabbro
Xu
X
Chen
Jacqueline E
JE
Baumgartner
Heidi A
HA
Lane
James A
JA
Fagin
D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000235:genetics*; D002460:Cell Line; D017931:DNA Primers; D066246:ErbB Receptors / Q000378:metabolism*; D047468:Immunoprecipitation; D051379:Mice; D011485:Protein Binding; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics* / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000098706:Taste Receptors, Type 2
D017931:DNA Primers; D043562:Receptors, G-Protein-Coupled; D000098706:Taste Receptors, Type 2; D066246:ErbB Receptors; D051096:Proto-Oncogene Proteins c-ret; C099282:RET protein, human