| Autor: |
Formento, J. L., Berra, E., Ferrua, B., Magne´, N., Simos, G., Brahimi-Horn, C., Pouysse´gur, J., Milano, G. |
| Zdroj: |
Clinical and Vaccine Immunology (formerly CDLI); May 2005, Vol. 12 Issue: 5 p660-664, 5p |
| Abstrakt: |
ABSTRACTHypoxia-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-1a and HIF-1ß. The only analytical methods available for measuring HIF-1a levels in tumors are immunohistochemistry and Western blotting. Immunohistochemistry has the advantage of allowing the identification and direct examination of HIF-1a-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-1a levels in cultured tumor cell lines in vitro. HIF-1a 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-1a under hypoxic conditions. The ELISA developed in the present study is thus an interesting alternative to other analytical methods used to measure HIF-1a protein levels and should be useful in preclinical pharmacological studies targeting HIF-1a. |
| Databáze: |
Supplemental Index |
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