title: The impact of stress on tumor growth: peripheral CRF mediates tumor-promoting effects of stress.
abstract: BACKGROUND
  Stress has been shown to be a tumor promoting factor. Both clinical and laboratory studies have shown that chronic stress is associated with tumor growth in several types of cancer. Corticotropin Releasing Factor (CRF) is the major hypothalamic mediator of stress, but is also expressed in peripheral tissues. Earlier studies have shown that peripheral CRF affects breast cancer cell proliferation and motility. The aim of the present study was to assess the significance of peripheral CRF on tumor growth as a mediator of the response to stress in vivo.
  
  
  METHODS
  For this purpose we used the 4T1 breast cancer cell line in cell culture and in vivo. Cells were treated with CRF in culture and gene specific arrays were performed to identify genes directly affected by CRF and involved in breast cancer cell growth. To assess the impact of peripheral CRF as a stress mediator in tumor growth, Balb/c mice were orthotopically injected with 4T1 cells in the mammary fat pad to induce breast tumors. Mice were subjected to repetitive immobilization stress as a model of chronic stress. To inhibit the action of CRF, the CRF antagonist antalarmin was injected intraperitoneally. Breast tissue samples were histologically analyzed and assessed for neoangiogenesis.
  
  
  RESULTS
  Array analysis revealed among other genes that CRF induced the expression of SMAD2 and β-catenin, genes involved in breast cancer cell proliferation and cytoskeletal changes associated with metastasis. Cell transfection and luciferase assays confirmed the role of CRF in WNT- β-catenin signaling. CRF induced 4T1 cell proliferation and augmented the TGF-β action on proliferation confirming its impact on TGFβ/SMAD2 signaling. In addition, CRF promoted actin reorganization and cell migration, suggesting a direct tumor-promoting action. Chronic stress augmented tumor growth in 4T1 breast tumor bearing mice and peripheral administration of the CRF antagonist antalarmin suppressed this effect. Moreover, antalarmin suppressed neoangiogenesis in 4T1 tumors in vivo.
  
  
  CONCLUSIONS
  This is the first report demonstrating that peripheral CRF, at least in part, mediates the tumor-promoting effects of stress and implicates CRF in SMAD2 and β-catenin expression.
authors.affiliation: Department of Clinical Chemistry, School Of Medicine, University of Crete, 71003 Heraklion, Greece.
authors.forename: Alicia
authors.initials: A
authors.lastname: Arranz
authors.forename: Maria
authors.initials: M
authors.lastname: Venihaki
authors.forename: Berber
authors.initials: B
authors.lastname: Mol
authors.forename: Ariadne
authors.initials: A
authors.lastname: Androulidaki
authors.forename: Erini
authors.initials: E
authors.lastname: Dermitzaki
authors.forename: Olga
authors.initials: O
authors.lastname: Rassouli
authors.forename: Jorge
authors.initials: J
authors.lastname: Ripoll
authors.forename: Efstathios N
authors.initials: EN
authors.lastname: Stathopoulos
authors.forename: Rosa P
authors.initials: RP
authors.lastname: Gomariz
authors.forename: Andrew N
authors.initials: AN
authors.lastname: Margioris
authors.forename: Christos
authors.initials: C
authors.lastname: Tsatsanis
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000209:etiology / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D003346:Corticotropin-Releasing Hormone / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011743:Pyrimidines / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology; D011863:Radioimmunoassay; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013312:Stress, Physiological / Q000235:genetics / Q000502:physiology*
chemical_list: D011743:Pyrimidines; D011758:Pyrroles; C103016:antalarmin; D003346:Corticotropin-Releasing Hormone