Hypoxia

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.



Principle

SIRION`s reporter system is based on real time bioluminescence imaging of HIF-1α activity. A luciferase protein is induced through the HRE (hypoxia responsive element of HIF-1α) under hypoxic conditions and targeted to proteasomal degradation under normoxia.


SIRION`s benefits


  • 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

Application Example


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.



Luciferase activity was determined using „Steady Glow Luciferase Assay“ from Promega


Service Details


SIRION offers custom generation of any desired hypoxia cell model making use of its unique reporter system. Figure out our custom cell model service or contact us for detailed project discussion.