Hypoxic microenvironment is coincidental with development and maintenance of tumors. The deeper understanding of how hypoxia affects angiogenesis and metabolism is therefore a research field of highest priority.
SIRION`SenCELL HypoxiaTM reporter system enables real time bioluminescence imaging of hypoxia in any in vitro cell or xenograft model. It exploits the natural characteristic of hypoxia and the hypoxia induced activation of the bHLH transcription factor HIF-1α (hypoxia inducible transcription factor-1α) .
Molecular charcteristics of HIF-1α
A) Under normoxia HIF-1α interacts with VHL (von Hippel-Lindau tumor suppressor protein) which leads to ubiquitination and subsequent proteasomal degradation.
B) Under hypoxia VHL cannot bind to HIF-1α leading to stabilization and activation.
- Very short half-life reporter protein (approx. 6min) → fast dynamic readout possible
- rapid response to hypoxia mimetics, proteasome inhibitors, prolyl hydroxylase inhi-bitors
- suitable e.g. for PD studies of anti-angiogenics
Generation of a stable HRE-Luciferase colon cancer reporter cell line by lentiviral transduction.
Hypoxia was induced using CoCl2 as mimetic agent occupying the VHL binding domain. Chemoluminescence is highly induced after CoCl2 treatment in HRE-Luciferase overexpressing cell line HRE-Luc2.