title: HIF-1A, pimonidazole, and iododeoxyuridine to estimate hypoxia and perfusion in human head-and-neck tumors.
abstract: OBJECTIVE
  Tumor hypoxia measured by microelectrodes has been shown to indicate poor patient outcome. Here we investigated four potentially more widely applicable immunohistochemical parameters of tumor oxygenation and perfusion in human head-and-neck tumors.
  
  
  METHODS
  Twenty patients with squamous cell carcinomas of the head and neck treated with primary surgery were injected with pimonidazole and IdUrd the evening before operation. Consecutive paraffin-embedded sections were stained for blood vessels, pimonidazole, IdUrd, and HIF-1alpha. IdUrd labeling and Ki-67 labeling around individual blood vessels were scored. The spatial relationship between HIF-1alpha and pimonidazole was studied, as well as the distribution of both markers as a function of distance from the nearest blood vessel.
  
  
  RESULTS
  Measurement of all four parameters (diffusion-limited fraction, pimonidazole fraction, HIF-1alpha fraction, IdUrd-negative vessels) was feasible, and a significant difference between tumors was found for all parameters. IdUrd-labeled cells were absent around some vessels, indicating lack of perfusion, because these regions were positive for Ki-67. There was a positive correlation between diffusion-limited fraction and pimonidazole area for all images from all tumors, although no correlation for mean values per tumor. Colocalization of pimonidazole and HIF-1alpha was low (0.02%-25%). Most expression profiles showed a more homogenous distribution for HIF-1alpha than pimonidazole. There was no significant correlation between the pimonidazole and HIF-1alpha fractions in the 10 tumors studied.
  
  
  CONCLUSIONS
  Simultaneous immunohistochemical measurements related to hypoxia and perfusion are feasible (and easily applicable) in resected human tumors. The different geographic distributions of HIF-1alpha and pimonidazole indicate that HIF-1alpha might not be suitable as a marker for chronic hypoxia. Each parameter will be correlated with outcome in a larger ongoing study on head-and-neck tumors treated with surgery with or without postoperative radiotherapy.
authors.affiliation: Division of Experimental Therapy, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors.forename: Hilde L K
authors.initials: HL
authors.lastname: Janssen
authors.forename: Karin M G
authors.initials: KM
authors.lastname: Haustermans
authors.forename: Debbie
authors.initials: D
authors.lastname: Sprong
authors.forename: Gerard
authors.initials: G
authors.lastname: Blommestijn
authors.forename: Ingrid
authors.initials: I
authors.lastname: Hofland
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Hoebers
authors.forename: Elke
authors.initials: E
authors.lastname: Blijweert
authors.forename: James A
authors.initials: JA
authors.lastname: Raleigh
authors.forename: Gregg L
authors.initials: GL
authors.lastname: Semenza
authors.forename: Mahesh A
authors.initials: MA
authors.lastname: Varia
authors.forename: Alfons J
authors.initials: AJ
authors.lastname: Balm
authors.forename: Marie Louise F
authors.initials: ML
authors.lastname: van Velthuysen
authors.forename: Pierre
authors.initials: P
authors.lastname: Delaere
authors.forename: Raf
authors.initials: R
authors.lastname: Sciot
authors.forename: Adrian C
authors.initials: AC
authors.lastname: Begg
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000175:diagnosis / Q000473:pathology; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D006258:Head and Neck Neoplasms / Q000175:diagnosis* / Q000473:pathology*; D006801:Humans; D000860:Hypoxia; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007065:Idoxuridine / Q000378:metabolism*; D007150:Immunohistochemistry; D019394:Ki-67 Antigen / Q000096:biosynthesis; D009389:Neovascularization, Pathologic; D009593:Nitroimidazoles / Q000378:metabolism*; D019384:Nucleic Acid Synthesis Inhibitors / Q000378:metabolism; D010477:Perfusion; D014157:Transcription Factors / Q000096:biosynthesis*; D016896:Treatment Outcome
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D019394:Ki-67 Antigen; D009593:Nitroimidazoles; D019384:Nucleic Acid Synthesis Inhibitors; D014157:Transcription Factors; C033815:pimonidazole; D007065:Idoxuridine