HIF-1A, pimonidazole, and iododeoxyuridine to estimate hypoxia and perfusion in human head-and-neck tumors.
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.
Janssen
Hilde L K
HL
Division of Experimental Therapy, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Haustermans
Karin M G
KM
Sprong
Debbie
D
Blommestijn
Gerard
G
Hofland
Ingrid
I
Hoebers
Frank J
FJ
Blijweert
Elke
E
Raleigh
James A
JA
Semenza
Gregg L
GL
Varia
Mahesh A
MA
Balm
Alfons J
AJ
van Velthuysen
Marie Louise F
ML
Delaere
Pierre
P
Sciot
Raf
R
Begg
Adrian C
AC
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
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