ABSTRACT
Erythropoietin (EPO) stimulates the growth of erythroblasts in the bone marrow (C. Lacombe and P. Mayeux, NEPHROL: DIAL: TRANSPLANT:, 14 (SUPPL: 2): 22-28, 1999). We report basal and hypoxia-stimulated expression of EPO and its receptor, EPOR, in human breast cancer cells, and we demonstrate EPO-stimulated tyrosine phosphorylation and the proliferation of these cells in vitro. In 50 clinical specimens of breast carcinoma, we report high levels of EPO and EPOR associated with malignant cells and tumor vasculature but not with normal breast, benign papilloma, or fibrocystic tissue. Hypoxic tumor regions display the highest levels of EPO and EPOR expression. Enhanced EPO signaling may contribute to the promotion of human cancer by tissue hypoxia.
Subject(s)
Breast Neoplasms/metabolism , Erythropoietin/biosynthesis , Receptors, Erythropoietin/biosynthesis , Biopsy , Blotting, Western , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Erythropoietin/genetics , Gene Expression , Humans , Immunohistochemistry , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptors, Erythropoietin/genetics , Reverse Transcriptase Polymerase Chain Reaction , Tumor Cells, CulturedABSTRACT
The cytokine ciliary neurotrophic factor (CNTF) and transforming growth factor-beta (TGF-beta) both induce transcription of the vasoactive intestinal peptide (VIP) gene through a 180-base pair cytokine response element (CyRE) in the VIP promoter. While CNTF induces STAT and AP-1 proteins to bind to cognate sites in the VIP CyRE, the mechanism through which TGF-beta acts to induce VIP gene transcription is not known. Here we show that Smad3 and Smad4 proteins can bind to two distinct sites within the VIP CyRE. These sites are absolutely required for the induction of VIP CyRE transcription by TGF-beta. TGF-beta induces endogenous Smad-containing complexes to bind to these sites in human neuroblastoma cells. CNTF and TGF-beta synergize to induce VIP mRNA expression and transcription through the VIP CyRE. This synergy is dependent on the Smad, STAT, and AP-1 sites, suggesting that these two independent cytokine pathways synergize through the cooperation of pathway-specific transcription factors binding to distinct sites within the VIP CyRE.
Subject(s)
Ciliary Neurotrophic Factor/physiology , Gene Expression Regulation/physiology , Transcription Factors/physiology , Transcription, Genetic/physiology , Transforming Growth Factor beta/physiology , Vasoactive Intestinal Peptide/genetics , Animals , Base Sequence , Cell Line , DNA PrimersABSTRACT
Activin, a member of the transforming growth factor-beta superfamily, can regulate neuropeptide gene expression in the nervous system and in neuroblastoma cells. Among the neuropeptide genes whose expression can be regulated by activin is the gene encoding the neuropeptide vasoactive intestinal peptide (VIP). To investigate the molecular mechanisms by which activin regulates neuronal gene expression, we have examined activin's regulation of VIP gene expression in NBFL neuroblastoma cells. We report here that NBFL cells respond to activin by increasing expression of VIP mRNA. Activin regulates VIP gene transcription in NBFL cells through a 180-bp element in the VIP promoter that was previously characterized to be necessary and sufficient to mediate the induction of VIP by the neuropoietic cytokines and termed the cytokine response element (CyRE). We find that the VIP CyRE is necessary and sufficient to mediate the transcriptional response to activin. In addition, ciliary neurotrophic factor (CNTF), a neuropoietic cytokine, synergizes with activin to increase VIP mRNA expression and transcription through the VIP CyRE. Mutations in either the Stat (signal transducer and activator of transcription) or AP-1 sites within the CyRE that reduce the response to CNTF, also reduce the response to activin. However, mutating both the Stat and AP-1 sites within the wild-type CyRE, while reducing the separate responses to either activin or CNTF, eliminates the synergy between them. These data suggest that activin and CNTF, two factors that appear to signal though distinct pathways, activate VIP gene transcription through a common transcriptional element, the VIP CyRE.
Subject(s)
Ciliary Neurotrophic Factor/pharmacology , Gene Expression Regulation/drug effects , Inhibins/pharmacology , Regulatory Sequences, Nucleic Acid , Signal Transduction/drug effects , Vasoactive Intestinal Peptide/biosynthesis , Activins , Animals , Binding Sites/genetics , Chickens , DNA-Binding Proteins/metabolism , Drug Synergism , Humans , Mutation , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics , Neuroblastoma/metabolism , Neuroblastoma/pathology , Promoter Regions, Genetic , Recombinant Fusion Proteins/biosynthesis , STAT1 Transcription Factor , Trans-Activators/metabolism , Transcription Factor AP-1/metabolism , Transfection , Transforming Growth Factor beta/pharmacology , Tumor Cells, Cultured/drug effects , Vasoactive Intestinal Peptide/geneticsABSTRACT
Eighteen men aged 18 to 32 with rapidly progressive male pattern baldness had serum dehydroepiandrosterone sulfate and testosterone measured. Dehydroepiandrosterone sulfate levels were elevated in all patients, ranging from 340 to 730 micrograms/dl. The patients were otherwise healthy and serum testosterone levels were within normal limits. A control group of men of similar age without hair loss had lower dehydroepiandrosterone sulfate levels ranging from 124 to 300 micrograms/dl (p less than 0.005). The biochemistry of androgens, particularly dehydroepiandrosterone sulfate, suggests that adrenal hyperactivity may initiate alopecia in young men who are genetically susceptible.
