Nuclear Translocation Of HIF-1α Research Articles

Butyrate impairs intestinal tumor cell induced angiogenesis by inhibiting HIF-1α nuclear translocation

increased c-Jun dephosphorylation was not involved. However, an inhibitor of tyrosinespecific phosphatases, orthovanadate, did restore activation-induced phosphorylation of cJun and cytokine production. A correlation ofc-Jun phosphorylation and cytokine production was also seen in T cells derived from cancer patients. Conclusion: Oxidative stress mimicked by hydrogen peroxide treatment inhibited T cell function. This inhibition is at least in part due to a reduction in c-Jun phosphorylation. This effect is mediated by tyrosine phospatase activity and possibly effects upstreamJNK, which requires phosphorylation of its threonin and tyrosine residues to phosphorylate c-Jun. A reduced susceptibility of c-Jun phosphorylation following stimulation was also observed in T cells derived from cancer patients. In accordance with our previous results this data suggests that the cellular immune response in cancer patients is influenced by the redox status of T cells and supports the concept of an antioxidant therapy or a treatment that reduces granulocyte activity.

Butyrate impairs intestinal tumor cell-induced angiogenesis by inhibiting HIF-1α nuclear translocation

Butyrate is known to stimulate proliferation of normal crypt cells, whereas it inhibits growth and induces apoptosis in colon cancer cells. We examined the effects of butyrate on colon cancer (Caco-2) cell-induced angiogenesis. HUVEC proliferation was significantly inhibited when incubated with medium conditioned by butyrate-treated Caco-2 cells. Simultaneously, levels of the proangiogenic vascular endothelial growth factor (VEGF) were reduced. HIF-1α protein, a transcription factor known to be a key regulator in hypoxia-induced angiogenesis, was upregulated by butyrate. This is in contrast to its importance as a VEGF regulating component. However Western blot of nuclear extracts revealed a downregulation of HIF-1α protein. HIF-1α DNA-binding activity was also decreased by butyrate. Our findings indicate that HIF-1α nuclear sequestration is repressed by butyrate, through inhibition of nuclear translocation. We postulate that diminished HIF-1α nuclear presence and activity in butyrate-treated Caco-2 cells could be responsible for decreased VEGF expression and antiangiogenic effects.

Epolones induce erythropoietin expression via hypoxia-inducible factor-1α activation

Induction of erythropoietin (Epo) expression under hypoxic conditions is mediated by the heterodimeric hypoxia-inducible factor (HIF)-1. Following binding to the 3' hypoxia-response element (HRE) of the Epo gene, HIF-1 markedly enhances Epo transcription. To facilitate the search for HIF-1 (ant)agonists, a hypoxia-reporter cell line (termed HRCHO5) was constructed containing a stably integrated luciferase gene under the control of triplicated heterologous HREs. Among various agents tested, we identified a class of substances called epolones, which induced HRE-dependent reporter gene activity in HRCHO5 cells. Epolones are fungal products known to induce Epo expression in hepatoma cells. We found that epolones (optimal concentration 4-8 micromol/L) potently induce HIF-1 alpha protein accumulation and nuclear translocation as well as HIF-1 DNA binding and reporter gene transactivation. Interestingly, the activity of a compound related to the fungal epolones, ciclopirox olamine (CPX), was blocked after addition of ferrous iron. This suggests that CPX might interfere with the putative heme oxygen sensor, as has been proposed for the iron chelator deferoxamine mesylate (DFX). However, about 10-fold higher concentrations of DFX (50-100 micromol/L) than CPX were required to maximally induce reporter gene activity in HRCHO5 cells. Moreover, structural, functional, and spectrophotometric data imply a chelator:iron stoichiometry of 1:1 for DFX but 3:1 for CPX. Because the iron concentration in the cell culture medium was determined to be 16 micromol/L, DFX but not CPX function can be explained by complete chelation of medium iron. These results suggest that the lipophilic epolones might induce HIF-1 alpha by intracellular iron chelation. (Blood. 2000;96:1558-1565)

Epolones induce erythropoietin expression via hypoxia-inducible factor-1α activation

AbstractInduction of erythropoietin (Epo) expression under hypoxic conditions is mediated by the heterodimeric hypoxia-inducible factor (HIF)-1. Following binding to the 3′ hypoxia-response element (HRE) of the Epo gene, HIF-1 markedly enhances Epo transcription. To facilitate the search for HIF-1 (ant)agonists, a hypoxia-reporter cell line (termed HRCHO5) was constructed containing a stably integrated luciferase gene under the control of triplicated heterologous HREs. Among various agents tested, we identified a class of substances called epolones, which induced HRE-dependent reporter gene activity in HRCHO5 cells. Epolones are fungal products known to induce Epo expression in hepatoma cells. We found that epolones (optimal concentration 4-8 μmol/L) potently induce HIF-1α protein accumulation and nuclear translocation as well as HIF-1 DNA binding and reporter gene transactivation. Interestingly, the activity of a compound related to the fungal epolones, ciclopirox olamine (CPX), was blocked after addition of ferrous iron. This suggests that CPX might interfere with the putative heme oxygen sensor, as has been proposed for the iron chelator deferoxamine mesylate (DFX). However, about 10-fold higher concentrations of DFX (50-100 μmol/L) than CPX were required to maximally induce reporter gene activity in HRCHO5 cells. Moreover, structural, functional, and spectrophotometric data imply a chelator:iron stoichiometry of 1:1 for DFX but 3:1 for CPX. Because the iron concentration in the cell culture medium was determined to be 16 μmol/L, DFX but not CPX function can be explained by complete chelation of medium iron. These results suggest that the lipophilic epolones might induce HIF-1α by intracellular iron chelation.