Hi Preethi,

In the study of Creig et al. of hemangioblastomas, the most frequent manifestation of hereditary von Hippel-Lindau disease (VHL), they found elevated levels of vascular endothelial growth factor and HIF-2  mRNA in stromal cells of the tumors. Mutations of the VHL tumor suppressor gene are associated with a variety of tumors such as renal clear cell carcinomas (RCC). Thereafter (please find a citation beneath), they analyzed the expression of the hypoxia-inducible factor HIF-2  in a range of VHL wild-type and VHL deficient RCC cell lines. In the presence of functional VHL protein, HIF-2  mRNA expression is increased only in cells lacking a functional VHL gene product. On the protein levels, however, in VHL deficient cell lines, HIF-2 was constitutively expressed, whereas re-introduction of a functional VHL gene restores the instability of HIF-2  proteins under normoxic conditions. Moreover, immunohistochemical analyses of RCCs and hemangioblastomas demonstrate up-regulation of HIF-2  in the tumor cells. The data presented provided evidence for a role of the VHL protein in regulation of angiogenesis and erythropoiesis mediated by the  HIF-2  protein. Oncogene (2000) 19, 5435-5443.

Best wishes,

Ilya

Hi llya,

Thank you very much for your reply so as of now there is no chemical available to induce HIF-2alpha.

Hi Preethi. You can try CoCl2, a common chemical compound that activates HIF-1alpha. 

Hi Poulios,

I am already working on that for HIF1 alpha I also want to induce HIF2 for an another study

Hypoxic treatment using a hypoxia chamber is probably the most physiological way to stabilize HIFs, other than that CoCl2 and MLN4924 (a NAE1 inhibitor, Soucy et al., Nature 2009) both stabilize HIFs in its non-ubiquitilated form, whereas proteasome inhibitors like bortezomib, carfilzomib or MG132 stabilize HIFs in its ubiquitilated form. Genetically, knockdown or knockout of VHL leads to HIF stabilization, if you want to create a cell line that stably accumulates a pool of HIFs. By HIFs, I am referring to HIF1alpha and HIF2alpha, since they are both following similar degradation pathways. 

Hai Kurt,

Thank you very much for your reply. I am already working on HIF-1alpha with both chemical as well as physiological induction but looking something specific for HIF-2alpha. Thank you very much for your valuable inputs.

Dear Preethi,

A recent paper indicate that hypoxia+acetate could induce acetilation of HIF-2 in hepatocyte (Chen et al, 2015 PLoS One). Moreover, Inoue et al (also in 2015) indicate that hydrostatic pressure can induce HIF-2 expression. 

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4331492/

http://www.mdpi.com/1422-0067/16/1/1043/pdf

Another way to look at HIF2alpha only is the 786-0 cell line, a RCC cell line that only expresses HIF2alpha but not HIF1alpha and also doesnt express VHL. I used those cells and they stably express HIF2alpha, I also made them stably expressing VHL which drives the levels down below detection and then the system reacts again to stabilizing reagents like CoCl2 or MLN or hypoxia. Maybe that cell line is an option for you...