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Clinical and Diagnostic Laboratory Immunology, May 2005, p. 660-664, Vol. 12, No. 5
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.5.660-664.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Enzyme-Linked Immunosorbent Assay for Pharmacological Studies Targeting Hypoxia-Inducible Factor 1{alpha}

J. L. Formento,1* E. Berra,2 B. Ferrua,3 N. Magné,1 G. Simos,4 C. Brahimi-Horn,2 J. Pouysségur,2 and G. Milano1

Oncopharmacology Unit of the Centre Antoine Lacassagne,1 Institute of Signaling, Developmental Biology and Cancer Research, CNRS UMR 6543,2 Medical School, Nice, France,3 University of Thessaly, Larissa, Greece4

Received 2 December 2004/ Returned for modification 3 January 2005/ Accepted 23 February 2005

Hypoxia-inducible factor 1 (HIF-1) activates the transcription of a wide range of genes related to oxygen delivery and metabolic adaptation under hypoxic (low-oxygen) conditions. HIF-1 is, in fact, a heterodimer of two subunits, HIF-1{alpha} and HIF-1ß. The only analytical methods available for measuring HIF-1{alpha} levels in tumors are immunohistochemistry and Western blotting. Immunohistochemistry has the advantage of allowing the identification and direct examination of HIF-1{alpha}-expressing cells, but has the intrinsic limitation, as for Western blotting, of being nonquantitative. We developed and validated an enzyme-linked immunosorbent assay (ELISA) approach to measure HIF-1{alpha} levels in cultured tumor cell lines in vitro. HIF-1{alpha} was expressed in thirteen tumor cell lines grown under hypoxic conditions; however, the levels differed strongly between cell lines. These data point to intrinsic differences between cell lines for the induction of HIF-1{alpha} under hypoxic conditions. The ELISA developed in the present study is thus an interesting alternative to other analytical methods used to measure HIF-1{alpha} protein levels and should be useful in preclinical pharmacological studies targeting HIF-1{alpha}.

* Corresponding author. Mailing address: Oncopharmacology Unit Centre, Antoine Lacassagne 33, Avenue de Valombrose, 06189 Nice Cedex 2, France. Phone: 33.(4).92.03.15.53. Fax: 33.(4).93.81.71.31. E-mail: jean-louis.formento{at}nice.fnclcc.fr.

Clinical and Diagnostic Laboratory Immunology, May 2005, p. 660-664, Vol. 12, No. 5
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.5.660-664.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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