i.e. antibody staining applied on FFPE sections of tumor tissue that reveals gradient positivity correlating to O2 tension?
my first suggestion would be, to stain for hypoxia-induced factor. But you probably already thought this over and rejected this as indirect marker.
Hello
HIF1α, HIF2α, GLUT1, GLUT4 and CAIX
http://jhc.sagepub.com/content/52/6/837.full
http://www.ncbi.nlm.nih.gov/pubmed/11121659
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3136703
Hello Fredrik,
PIMO is not reliable. It is OK for ex-vivo experiments but that is all.
Applied intravenously on mouse models it can be fine too but it will not penetrate far from the vessels and that is where hypoxia is established.
HIF family is extremely unstable and also can be upregulated during the cold ischemia tissue "suffers" after resection.
Colateral damage molecules like aldehydes as a result of lipid peroxydation, might be a stable hallmark of hypoxic tissues and are relatively stable even in FFPE tissues.
I used anti 4-hydroxynonenal antibody which worked fine. The IHC was in line with the EPR data, so more or less it is concomitant to hypoxia.
Another option is upregulation of some NADPH oxidases, which are primary target genes of HIF1 alpha for example or prolylhydroxylase (PHD) IHC, as they will also be upregulated and have direct relation to HIF degradation.
Cheers,
Ivan
Thanks for all answers! The anti 4-hydroxynonenal antibody sounds most interesting, any nice image examples of hypoxic regions/zones in tumor tissue? Trying to look through some of the cancer tissues in the Human Protein Atlas (proteinatlas.org) to find patterns that show some resemblance to what could be grade of hypoxia.......
Hi Fredrik,
That is where in cancer things become "creepy". Due to the resection, followed by focal ischemia, prolonged fixation etc., one can mimic real hypoxia very nicely.
It is tough matter, I spent several years in the field before coming back to pathology and is quite. Pathology won, though :)
Best and write us should you need anything.
Best,
Ivan
Dear Dr.
i hop you get use of the following link from British Journal of Cancer (2015) 112, 382–390. doi:10.1038/bjc.2014.604
and also this link http://www.nature.com/bjc/journal/v112/n2/full/bjc2014604a.html
Take care when using proteins/genes from HIF pathway to check hypoxia in cancer, a lot of them will present altered pathway. HIF role in cancer is still under discussion. Anti 4-hydroxynonenal antibody will be a nice option.
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