Echinomycin, a Small-Molecule Inhibitor of Hypoxia-Inducible Factor-1 DNA-Binding Activity
Dehe Kong,Eun-Jung Park,Andrew G. Stephen,Maura Calvani,John H. Cardellina,Anne Monks,Robert J. Fisher,Robert H. Shoemaker,Giovanni Melillo +8 more
TLDR
It is shown that it is possible to identify small molecules that inhibit HIF-1 DNA binding to endogenous promoters and echinomycin, a small-molecule known to bind DNA in a sequence-specific fashion, was investigated.Abstract:
The identification of small molecules that inhibit the sequence-specific binding of transcription factors to DNA is an attractive approach for regulation of gene expression. Hypoxia-inducible factor-1 (HIF-1) is a transcription factor that controls genes involved in glycolysis, angiogenesis, migration, and invasion, all of which are important for tumor progression and metastasis. To identify inhibitors of HIF-1 DNA-binding activity, we expressed truncated HIF-1α and HIF-1β proteins containing the basic-helix-loop-helix and PAS domains. Expressed recombinant HIF-1α and HIF-1β proteins induced a specific DNA-binding activity to a double-stranded oligonucleotide containing a canonical hypoxia-responsive element (HRE). One hundred twenty-eight compounds previously identified in a HIF-1–targeted cell-based high-throughput screen of the National Cancer Institute 140,000 small-molecule library were tested in a 96-well plate ELISA for inhibition of HIF-1 DNA-binding activity. One of the most potent compounds identified, echinomycin (NSC-13502), a small-molecule known to bind DNA in a sequence-specific fashion, was further investigated. Electrophoretic mobility shift assay experiments showed that NSC-13502 inhibited binding of HIF-1α and HIF-1β proteins to a HRE sequence but not binding of the corresponding proteins to activator protein-1 (AP-1) or nuclear factor-κB (NF-κB) consensus sequences. Interestingly, chromatin immunoprecipitation experiments showed that NSC-13502 specifically inhibited binding of HIF-1 to the HRE sequence contained in the vascular endothelial growth factor (VEGF) promoter but not binding of AP-1 or NF-κB to promoter regions of corresponding target genes. Accordingly, NSC-13502 inhibited hypoxic induction of luciferase in U251-HRE cells and VEGF mRNA expression in U251 cells. Our results indicate that it is possible to identify small molecules that inhibit HIF-1 DNA binding to endogenous promoters.read more
Citations
More filters
Journal ArticleDOI
Tumor Cell Metabolism: Cancer's Achilles' Heel
TL;DR: The peculiarities of tumor cell metabolism are reviewed to discuss the alterations in signal transduction pathways and/or enzymatic machineries that account for metabolic reprogramming of transformed cells.
Journal ArticleDOI
Hypoxia signalling in cancer and approaches to enforce tumour regression
TL;DR: There are new approaches to enforce necrotic cell death and tumour regression by targeting tumour metabolism and pHi-control systems.
Journal ArticleDOI
Defining the role of hypoxia-inducible factor 1 in cancer biology and therapeutics
TL;DR: This review summarizes the current state of knowledge regarding the molecular mechanisms by which Hif-1 contributes to cancer progression, focusing on clinical data associating increased HIF-1 levels with patient mortality and pharmacological data showing anticancer effects of H IF-1 inhibitors in mouse models of human cancer.
Journal ArticleDOI
HIF-1α pathway: role, regulation and intervention for cancer therapy
Georgina N. Masoud,Wei Li +1 more
TL;DR: The role and regulation of the HIF-1α in cancer, and recent therapeutic approaches targeting this important pathway are summarized.
Journal ArticleDOI
Energy metabolism in tumor cells
TL;DR: It is proposed that energy metabolism may be an alternative therapeutic target for both hypoxic (glycolytic) and oxidative tumors.
References
More filters
Journal ArticleDOI
Targeting HIF-1 for cancer therapy
TL;DR: Hypoxia-inducible factor 1 (HIF-1) activates the transcription of genes that are involved in crucial aspects of cancer biology, including angiogenesis, cell survival, glucose metabolism and invasion.
Journal ArticleDOI
Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension
TL;DR: Hypoxia-inducible factor 1 (HIF-1) is found in mammalian cells cultured under reduced O2 tension and is necessary for transcriptional activation mediated by the erythropoietin gene enhancer in hypoxic cells.
Journal ArticleDOI
The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis
Patrick H. Maxwell,Michael S. Wiesener,Gin-Wen Chang,Steven C. Clifford,Emma C. Vaux,Matthew Edward Cockman,Charles C. Wykoff,Christopher W. Pugh,Eamonn R. Maher,Peter J. Ratcliffe,Peter J. Ratcliffe +10 more
TL;DR: It is indicated that the interaction between HIF-1 and pVHL is iron dependent, and that it is necessary for the oxygen-dependent degradation of HIF α-subunits, which may underlie the angiogenic phenotype of VHL-associated tumours.
Journal ArticleDOI
Feasibility of a High-Flux Anticancer Drug Screen Using a Diverse Panel of Cultured Human Tumor Cell Lines
Anne Monks,Dominic A. Scudiero,Philip Skehan,Robert H. Shoemaker,Kenneth D. Paull,David T. Vistica,Curtis Hose,John Langley,Paul Cronise,Anne Vaigro-Wolff,Marcia Gray-Goodrich,H. D. Campbell,Joseph G. Mayo,Michael R. Boyd +13 more
TL;DR: A pilot-scale, in vitro, anticancer drug screen utilizing a panel of 60 human tumor cell lines organized into subpanels representing leukemia, melanoma, and cancers of the lung, colon, kidney, ovary, and central nervous system is described.
Journal Article
Overexpression of Hypoxia-inducible Factor 1α in Common Human Cancers and Their Metastases
Hua Zhong,Angelo M. De Marzo,Erik Laughner,Michael Lim,David A Hilton,David Zagzag,Peter Buechler,William B. Isaacs,Gregg L. Semenza,Jonathan W. Simons +9 more
TL;DR: The first clinical data indicating that HIF-1alpha may play an important role in human cancer progression are provided, indicating adaptations to a hypoxic microenvironment that are correlated with tumor invasion, metastasis, and lethality.