15-Deoxy-Δ12,14-prostaglandin J2stabilizes hypoxia inducible factor-1α through induction of heme oxygenase-1 and direct modification ofprolyl-4-hydroxylase 2.
15-Deoxy-Δ12,14-prostaglandin J2(15d-PGJ2), a representative J-series cyclopentenone prostaglandin, has biphasic roles in cell proliferation and apoptosis. Hypoxia inducible factor-1 (HIF-1) regulates expression of various genes involved in tumor growth and angiogenesis. In the present study, treatment of human breast cancer (MCF-7) cells with 15d-PGJ2resulted in the accumulation of the α-subunit of HIF-1. Pretreatment with zinc protoporphyrin IX, a pharmacological inhibitor of heme oxygenase-1 (HO-1), as well as siRNA knockdown of HO-1 gene in MCF-7 cells attenuated 15d-PGJ2-mediated HIF-1α accumulation. 15d-PGJ2treatment increased intracellular production of reactive oxygen species (ROS), which was mediated by HO-1 induction. Preincubation of MCF-7 cells with trolox, a water-soluble form of vitamin E, attenuated 15d-PGJ2-induced HIF-1α expression although HO-1 expression was unchanged. This finding suggests that ROS accumulated as a consequence of HO-1 up-regulation can enhance HIF-1α expression in MCF-7 cells treated with 15d-PGJ2. Alternatively, 15d-PGJ2was found to covalently bind to HIF-1α prolyl-4-hydroxylase 2 (PHD2) in MCF-7 cells, which hampers the proline hydroxylation of HIF-1α, thereby disrupting ubiquitin-dependent proteasomal degradation of this transcription factor. Pretreatment with thiol reducing agents blunted 15d-PGJ2-induced HIF-1α stabilization, indicative of a cysteine residue as a direct target of 15d-PGJ2. Molecular docking analysis suggests that 15d-PGJ2preferentially binds to PHD2 in the vicinity of the Cys201residue based on binding energies and carbon-sulfur distances. In summary, 15d-PGJ2stabilizes HIF-1α in MCF-7 cells through HO-1 induction with subsequent ROS generation and also through direct modification of PHD2.
a Research Institute of Pharmaceutical Sciences , College of Pharmacy , Seoul , South Korea.
Jee-Eun
JE
Choi
a Research Institute of Pharmaceutical Sciences , College of Pharmacy , Seoul , South Korea.
Jung-Hyun
JH
Kim
a Research Institute of Pharmaceutical Sciences , College of Pharmacy , Seoul , South Korea.
Na-Young
NY
Song
a Research Institute of Pharmaceutical Sciences , College of Pharmacy , Seoul , South Korea.
Jinyoung
J
Suh
a Research Institute of Pharmaceutical Sciences , College of Pharmacy , Seoul , South Korea.
Do-Hee
DH
Kim
b Department of Molecular Medicine and Biopharmaceutical Sciences , Graduate School of Convergence Sciences and Technology , Seoul , South Korea.
Su-Jung
SJ
Kim
c Department of Food and Nutrition , Sungshin Women's University , Seoul , South Korea.
Hye-Kyung
HK
Na
d Hormel Institute, University of Minnesota , Austin , MN , USA.
Janos
J
Nadas
b Department of Molecular Medicine and Biopharmaceutical Sciences , Graduate School of Convergence Sciences and Technology , Seoul , South Korea.|d Hormel Institute, University of Minnesota , Austin , MN , USA.
Zigang
Z
Dong
e College of Medicine , Inha University , Incheon , South Korea.
Young-Nam
YN
Cha
a Research Institute of Pharmaceutical Sciences , College of Pharmacy , Seoul , South Korea.|b Department of Molecular Medicine and Biopharmaceutical Sciences , Graduate School of Convergence Sciences and Technology , Seoul , South Korea.|f Cancer Research Institute , Seoul National University , Seoul , South Korea.
Young-Joon
YJ
Surh
D051547:Heme Oxygenase-1 / Q000378:metabolism; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D064828:Prolyl Hydroxylases / Q000378:metabolism*
D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D064828:Prolyl Hydroxylases; D051547:Heme Oxygenase-1
15-Deoxy-Δ12,14-prostaglandin J2; MCF-7 cells; cyclopentenone prostaglandin; heme oxygenase-1; hypoxia inducible factor-1α; prolyl-4-hydroxylase 2