title: EGF-mediated regulation of IGFBP-3 determines esophageal epithelial cellular response to IGF-I.
abstract: IGF and EGF regulate various physiological and pathological processes. IGF binding protein (IGFBP)-3 regulates cell proliferation in IGF-dependent and -independent fashions. Recently, we identified IGFBP-3 as a novel EGF receptor (EGFR) downstream target molecule in primary and immortalized human esophageal epithelial cells, suggesting an interplay between the EGF and IGF signaling pathways. However, the regulatory mechanisms for IGFBP-3 expression and its functional role in esophageal cell proliferation remain to be elucidated. Herein, we report that IGFBP-3 mRNA and protein were induced upon growth factor deprivation in primary and immortalized human esophageal cells through mechanisms requiring p53-independent de novo mRNA transcription and protein synthesis. This occurred in the face of the activated phosphatidylinositol 3-OH-kinase (PI3K)/mammalian target of rapamycin (mTOR) pathway. Secreted IGFBP-3 neutralized IGFs and prevented IGF-I receptor (IGF-IR) activation. In contrast, EGF suppressed IGFBP-3 mRNA and protein expression through activation of MAPK in an EGFR-tyrosine kinase-dependent manner to restore the cellular response to IGF-I. When stably overexpressed, wild-type IGFBP-3 but not I56G/L80G/L81G (GGG) mutant IGFBP-3, which has a reduced affinity to IGFs, prevented IGF-I from activating IGF-IR and Akt as well as stimulating cell proliferation. However, unlike other cell types where IGFBP-3 exerts antiproliferative effects, neither wild-type nor GGG mutant IGFBP-3 alone affected cell proliferation or EGFR activity. These results indicate that IGF signaling is subject to negative regulation through IGFBP-3 and positive regulation by EGF, the latter of which suppresses IGFBP-3. This provides a platform for understanding the novel cross talk between EGF- and IGF-mediated pathways.
authors.affiliation: Gastroenterology Division, University of Pennsylvania, 415 Curie Blvd., Philadelphia, PA 19104, USA.
authors.forename: Munenori
authors.initials: M
authors.lastname: Takaoka
authors.forename: Caitlin E
authors.initials: CE
authors.lastname: Smith
authors.forename: Michael K
authors.initials: MK
authors.lastname: Mashiba
authors.forename: Takaomi
authors.initials: T
authors.lastname: Okawa
authors.forename: Claudia D
authors.initials: CD
authors.lastname: Andl
authors.forename: Wafik S
authors.initials: WS
authors.lastname: El-Deiry
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nakagawa
mesh_terms: D015153:Blotting, Western; D049109:Cell Proliferation; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000502:physiology*; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology*; D004947:Esophagus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016158:Genes, p53 / Q000235:genetics; D011905:Genes, ras / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000378:metabolism / Q000502:physiology*; D007334:Insulin-Like Growth Factor I / Q000502:physiology*; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013936:Thymidine / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: D018972:Insulin-Like Growth Factor Binding Protein 3; D004815:Epidermal Growth Factor; D007334:Insulin-Like Growth Factor I; D019869:Phosphatidylinositol 3-Kinases; D020928:Mitogen-Activated Protein Kinases; D013936:Thymidine