title: Expression of erythropoietin and its receptor increases in colonic neoplastic progression: the role of hypoxia in tumorigenesis.
abstract: BACKGROUND
  Tissue hypoxia is a characteristic patho-physiologic property of colorectal cancer. This process may also add to a therapeutic problem of solid tumor resistance to chemo- and radiation therapy. Erythropoietin (Epo) expression is induced by tissue hypoxia. Acting via its receptor (EpoR), Epo inhibits apoptosis of erythroid cells and has been shown to rescue neurons from hypoxic damage. Increased Epo and EpoR expression has been recently described in human breast, renal and cervical carcinoma. Given the characteristic tumor diathesis present in majority of colorectal cancers, we examined whether Epo signaling may play a role in colonic neoplastic progression.
  
  
  METHODS
  Expression of Epo and EpoR was examined using immunohistochemistry in 24 cases of primary colorectal and metastatic adenocarcinomas versus adenomas and normal colonic mucosa. Immunohistochemical stains were evaluated semiquantitatively based on a four-tiered scale. Based on the combination of extent and intensity of immunoreactivity, an immunostaining score (0-300) was determined for each sample. Expression of Epo and EpoR protein and mRNA was examined using Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR), respectively, in both normal colonic tissue and carcinoma specimens in five cases.
  
  
  RESULTS
  Epo expression was sequentially increased in normal colonic mucosa (8.3 ± 5.6, mean ± SEM), adenoma (26.4 ± 9.1), primary carcinoma (96.1 ± 12.8) and metastatic carcinoma (122 ± 51.3). EpoR expression was also sequentially increased in normal colonic mucosa (22.3 ± 11.8), adenoma (108.7 ± 24.2), primary carcinoma (178.7 ± 16.6) and metastatic carcinoma (220 ± 58.3) (P< 0.05 for all results). Epo and EpoR showed enhanced expression in the areas adjacent to ischemia/necrosis. Western blot and RT-PCR analysis revealed increased EpoR protein and mRNA levels in carcinoma compared to normal mucosal colon specimens. Focal stromal Epo and EpoR immunoreactivity was present in 10 and 12 cases, respectively.
  
  
  CONCLUSIONS
  The uniform increase in the expression of Epo and EpoR along the colonic neoplastic sequence and further increase in ischemic/necrotic areas indicates that the Epo signaling pathway is an important component in colon carcinogenesis including possible epithelial-stromal interactions.
authors.affiliation: Department of Pathology, Lafayette General Medical Center, Lafayette, LA, USA. zgombos@lgmc.com
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Gombos
authors.forename: Ludovit
authors.initials: L
authors.lastname: Danihel
authors.forename: Vanda
authors.initials: V
authors.lastname: Repiska
authors.forename: Geza
authors.initials: G
authors.lastname: Acs
authors.forename: Emma
authors.initials: E
authors.lastname: Furth
mesh_terms: D000230:Adenocarcinoma / Q000473:pathology* / Q000556:secondary; D000236:Adenoma / Q000473:pathology*; D015153:Blotting, Western; D003110:Colonic Neoplasms / Q000473:pathology* / Q000556:secondary; D004921:Erythropoietin / Q000096:biosynthesis*; D015870:Gene Expression; D006801:Humans; D000860:Hypoxia; D007150:Immunohistochemistry; D008853:Microscopy; D017467:Receptors, Erythropoietin / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction
chemical_list: C554270:EPO protein, human; D017467:Receptors, Erythropoietin; D004921:Erythropoietin