23 results on '"Boudouresque, F."'
Search Results
2. Identification of secondary structure in the 5′-untranslated region of the human adrenomedullin mRNA with implications for the regulation of mRNA translation
- Author
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Brenet, F, Dussault, N, Delfino, C, Boudouresque, F, Chinot, O, Martin, P-M, and Ouafik, L H
- Published
- 2006
- Full Text
- View/download PDF
3. Identification of secondary structure in the 5'-untranslated region of the human adrenomedullin mRNA with implications for the regulation of mRNA translation
- Author
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L'Houcine Ouafik, Olivier Chinot, Fabienne Brenet, C. Delfino, Boudouresque F, Pierre-Marie Martin, Nadège Dussault, Ouafik, L'Houcine, Rôle et mécanismes d'action de l'adrenomedulline dans la croissance tumorale : Application au modèle des gliomes, Université de la Méditerranée - Aix-Marseille 2-IFR11-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherches en Oncologie biologique et Oncopharmacologie (CRO2), Institut National de la Santé et de la Recherche Médicale (INSERM)- Hôpital de la Timone [CHU - APHM] (TIMONE)-Aix Marseille Université (AMU), and Aix Marseille Université (AMU)- Hôpital de la Timone [CHU - APHM] (TIMONE)-Institut National de la Santé et de la Recherche Médicale (INSERM)
- Subjects
Untranslated region ,Cancer Research ,DNA, Complementary ,Five prime untranslated region ,Molecular Sequence Data ,RNA-binding protein ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Biology ,MESH: Base Sequence ,Adrenomedullin ,[SDV.CAN] Life Sciences [q-bio]/Cancer ,Genes, Reporter ,Genetics ,Humans ,Luciferase ,RNA, Messenger ,Molecular Biology ,Cells, Cultured ,MESH: RNA, Messenger ,Messenger RNA ,MESH: Humans ,MESH: Molecular Sequence Data ,Base Sequence ,MESH: Peptides ,MESH: Genes, Reporter ,RNA-Binding Proteins ,RNA ,Translation (biology) ,MESH: DNA, Complementary ,Stem-loop ,Molecular biology ,MESH: Adrenomedullin ,MESH: Sequence Alignm ,MESH: RNA-Binding Proteins ,MESH: Nucleic Acid Conformation ,Protein Biosynthesis ,MESH: Protein Biosynthesis ,Nucleic Acid Conformation ,MESH: 5' Untranslated Regions ,5' Untranslated Regions ,Peptides ,Sequence Alignment ,MESH: Cells, Cultured - Abstract
Adrenomedullin (AM) is a multifunctional regulatory peptide with important angiogenic and mitogenic properties. Here we identify a region of stable secondary structure in the 5'-untranslated region (5' UTR) of human AM mRNA. Reverse transcriptase-polymerase chain reaction of the 5' UTR consistently resulted, in addition to the product with the expected size of 155 base pair (bp), in a second product with an approximately 65-bp deletion from the central region of the 5' UTR, suggesting the presence of a secondary structure. The presence of a stem-loop structure was confirmed by probing the 5' UTR with RNases with selectivity for single- or double-stranded RNA. We investigated the role of this stem-loop structure in expression of luciferase reporter gene in cultured cell lines. Reporter assays using a chimeric mRNA that combined luciferase and the 5' UTR of AM mRNA demonstrated a dramatic decrease of the reporter activity owing to a decreased translation, whereas the deletion of the stem-loop structure localized between nt +31 and +95 from the cap site led to the recovery of activity. Gel migration shift assays using cytosolic extracts from mammalian cell lines demonstrate a specific binding of a cytosolic protein to riboprobes containing the 5' UTR of AM but not to riboprobes either corresponding to other areas of the message or containing the 5' UTR but lacking the region of secondary structure. Although we conclude that the 5' UTR of the human AM mRNA can modulate the translation of AM mRNA in vivo, and that the predicted stem-loop structure is necessary for this inhibition, the functional consequences of the cis element-binding activity remain to be determined.
- Published
- 2006
4. P04.08 * PLASMA LEVELS AND TUMOR TISSUE RNA OF MMP2 AND MMP9 EXHIBIT SIMILAR DISTRIBUTION IN NEWLY DIAGNOSED AND RECURRENT GLIOBLASTOMA (GB)
- Author
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Tabouret, E., primary, Boudouresque, F., additional, Denicolai, E., additional, Barrie, M., additional, Boucard, C., additional, Metellus, P., additional, Autran, D., additional, Figarella-Branger, D., additional, Ouafik, L., additional, and Chinot, O., additional
- Published
- 2014
- Full Text
- View/download PDF
5. ASSOCIATION OF MATRIX METALLOPROTEINASE 2 (MMP2) BASELINE PLASMA LEVEL WITH RESPONSE AND SURVIVAL AND CHANGE OVERTIME IN PATIENTS TREATED WITH BEVACIZUMAB FOR RECURRENT HIGH GRADE GLIOMA
- Author
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Chinot, O.-L., primary, Boudouresque, F., additional, Barrie, M., additional, Matta, M., additional, Boucard, C., additional, Loundou, A., additional, Figarella-Branger, D., additional, Ouafik, L., additional, and Tabouret, E., additional
- Published
- 2014
- Full Text
- View/download PDF
6. Expression of adrenomedullin and peptide amidation activity in human prostate cancer and in human prostate cancer cell lines
- Author
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Rocchi, P., Boudouresque, F., Zamora, A. J., Muracciole, X., Lechevallier, E., Martin, P. -M, L’Houcine Ouafik, LABORATOIRE DE CANCEROLOGIE EXPERIMENTALE EA2671, Université de la Méditerranée - Aix-Marseille 2, Laboratoire de Transfert d'Oncologie Biologique [Hôpital Nord - APHM], Hôpital Nord [CHU - APHM]-Assistance Publique - Hôpitaux de Marseille (APHM)-Aix Marseille Université (AMU), Interactions cellulaires neuroendocriniennes (ICN), Université de la Méditerranée - Aix-Marseille 2-Centre National de la Recherche Scientifique (CNRS), Service de Radiothérapie, APHM, Hôpital de la Timone, Marseille, Hôpital de la Timone [CHU - APHM] (TIMONE), SERVICE D'UROLOGIE, Hôpital Salvator [CHU - APHM] (Hôpitaux Sud), Ouafik, L'Houcine, and Aix Marseille Université (AMU)-Assistance Publique - Hôpitaux de Marseille (APHM)- Hôpital Nord [CHU - APHM]
- Subjects
Male ,MESH: Neoplasm Proteins ,Neoplasms, Hormone-Dependent ,Transplantation, Heterologous ,Prostatic Hyperplasia ,Mice, Nude ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Adenocarcinoma ,urologic and male genital diseases ,MESH: Multienzyme Complexes ,MESH: Phosphopyruvate Hydratase ,Mixed Function Oxygenases ,Adrenomedullin ,Mice ,MESH: Prostatic Hyperplasia ,MESH: In Situ Hybridization ,[SDV.CAN] Life Sciences [q-bio]/Cancer ,Multienzyme Complexes ,MESH: Reverse Transcriptase Polymerase Chain Reaction ,Tumor Cells, Cultured ,MESH: Mice, Nude ,Animals ,Humans ,MESH: Animals ,RNA, Messenger ,MESH: Tumor Cells, Cultured ,MESH: Neoplasms, Hormone-Dependent ,MESH: Transplantation, Heterologous ,MESH: Mice ,In Situ Hybridization ,MESH: RNA, Messenger ,MESH: Humans ,Reverse Transcriptase Polymerase Chain Reaction ,MESH: Peptides ,MESH: Adenocarcinoma ,Prostatic Neoplasms ,MESH: Immunohistochemistry ,MESH: Mixed Function Oxygenases ,Immunohistochemistry ,MESH: Adrenomedullin ,MESH: Male ,Neoplasm Proteins ,Phosphopyruvate Hydratase ,MESH: Prostatic Neoplasms ,MESH: Cell Division ,Peptides ,Cell Division ,Neoplasm Transplantation ,MESH: Neoplasm Transplantation - Abstract
International audience; After therapeutic hormone deprivation, prostate cancer (CaP) cells often develop androgen-independent growth through not-well-defined mechanisms. The presence of neuroendocrine (NE) cells is often greater in prostate carcinoma than in normal prostate, and the frequency of NE cells correlates with tumor malignancy, loss of androgen sensitivity, increase of autocrine-paracrine activity, and poor prognosis. In some CaPs, neuropeptides have been previously implicated as growth factors. Peptidylglycine alpha-amidating monooxygenase (PAM) is the enzyme producing alpha-amidated bioactive peptides from their inactive glycine-extended precursors. In the present work, we demonstrate that androgen-independent PC-3 and DU145 cell lines, derived from human CaP, express PAM in vitro and in xenografts implanted in athymic nude mice, indicating that they are able to produce alpha-amidated peptides. Contrarily, barely detectable levels of PAM were found in the androgen-sensitive LNCaP cell line. We also show that whereas PC-3 and DU145 cells produce and secrete adrenomedullin (AM), a multifunctional amidated peptide, no expression was found in LNCaP cells. We further demonstrate that AM acts as a growth factor for DU145 cells, which suggests the existence of an autocrine loop mechanism that could potentially drive neoplastic growth. PAM mRNA levels were found to be 3-fold higher in prostate adenocarcinomas compared with that of human benign prostate hyperplasia (BPH) as demonstrated by real-time quantitative reverse transcription-PCR. The analysis of AM message expression in BPH and CaP (Gleason's score, 6-9) shows a clear distinction between benign and CaP. The expression was detected only in adenocarcinomas tissues with a marked increase in samples with a high Gleason's score. Immunocytochemically, AM was localized in the carcinomatous epithelial compartment. NE phenotype, assessed after the immunocytochemical localization of neuron-specific enolase (NSE), was found in both the epithelial and the stromal compartments of cancers; in BPH, only some spare basal cells were NSE-labeled. Cancer progression could be accelerated by peptides secreted by a population of cells capable of inducing androgen-independent tumoral growth via autocrine-paracrine mechanisms.
- Published
- 2001
7. Association of Matrix Metalloproteinase 2 (MMP2) Plasma Level with Response and Survival in Patients Treated with Bevacizumab for Recurrent High Grade Glioma
- Author
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Tabouret, E., primary, Boudouresque, F., additional, Barrié, M., additional, Matta, M., additional, Boucard, C., additional, Loundou, A., additional, Ouafik, M., additional, and Chinot, O., additional
- Published
- 2012
- Full Text
- View/download PDF
8. Adrenomedullin, an autocrine/paracrine factor induced by androgen withdrawal, stimulates ‘neuroendocrine phenotype’ in LNCaP prostate tumor cells
- Author
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Berenguer, C, primary, Boudouresque, F, additional, Dussert, C, additional, Daniel, L, additional, Muracciole, X, additional, Grino, M, additional, Rossi, D, additional, Mabrouk, K, additional, Figarella-Branger, D, additional, Martin, P-M, additional, and Ouafik, L'Houcine, additional
- Published
- 2007
- Full Text
- View/download PDF
9. Insulin-induced hypoglycemia stimulates corticotropin-releasing factor and arginine vasopressin secretion into hydrophysical portal blood of conscious, unrestrained rams
- Author
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Caraty, Alain, Grino, Michel, Locatelli, Aude, Guillaume, V., Boudouresque, F., Conte-Devolx, B., Oliver, Charlotte, ProdInra, Migration, Unité de recherche Physiologie de la reproduction des mammifères domestiques, Nouzilly, and Institut National de la Recherche Agronomique (INRA)
- Subjects
[SDV] Life Sciences [q-bio] ,ADRENOCORTICOTROPINE ,[SDV]Life Sciences [q-bio] ,CORTISOL ,[INFO]Computer Science [cs] ,CRF ,[INFO] Computer Science [cs] ,ComputingMilieux_MISCELLANEOUS - Abstract
International audience
- Published
- 1990
10. Effect of thyroid hormones on peptidylglycine alpha-amidating monooxygenase gene expression in anterior pituitary gland: transcriptional studies and messenger ribonucleic acid stability.
- Author
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Fraboulet, S, primary, Boudouresque, F, additional, Delfino, C, additional, Fina, F, additional, Oliver, C, additional, and Ouafik, L, additional
- Published
- 1996
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11. Evidence for high peptide alpha-amidating activity in the pancrease from neonatal rats.
- Author
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Ouafik, L, Giraud, P, Salers, P, Dutour, A, Castanas, E, Boudouresque, F, and Oliver, C
- Abstract
A high peptidylglycine alpha-amidating mono-oxygenase (PAMase) activity has been measured in the pancreas of neonatal rats. A significant fraction of this activity is contained in the beta cells of the islets of Langerhans and is colocalized with thyrotropin-releasing hormone (TRH) and its precursor in secretory granules. The ontogenetic variation of PAMase activity in the pancrease parallels that of TRH concentrations, suggesting that this enzymatic activity is directly related to TRH biosynthesis. In addition, PAMase activity is able to generate TRH when incubated with less than Glu-His-Pro-Gly, a tetrapeptide present as a repetitive sequence in the TRH precursor. The perinatal evolution of the TRH precursor levels in the pancreas is similar to that of PAMase activity (unpublished results). Thus, the neonatal rat pancreas offers an endocrine model in which the levels of a neuropeptide precursor and an enzyme activity, involved in the posttranslational modification of this precursor, are similarly regulated. Our results suggest also that a fraction of PAMase activity may be produced outside of the beta cells and related to the biosynthesis of COOH-terminally amidated peptide(s) other than TRH. The ontogenetic changes in PAMase activity imply that the synthesis of this peptide(s) is high during the neonatal period, decreasing thereafter.
- Published
- 1987
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12. Ribonuclease MCPiP1 contributes to the loss of micro-RNA-200 family members in pancreatic cancer cells.
- Author
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Boudouresque F, Siret C, Dobric A, Silvy F, Soubeyran P, Iovanna J, Lombardo D, and Berthois Y
- Abstract
The microRNA-200 (miR-200) family is frequently down-regulated in tumors, including pancreatic adenocarcinomas (PDACs). In this study we have examined the mechanisms involved in the loss of miR-200s in tumoral pancreatic cells. Whereas miR-200 gene promoters appear methylated in mature miR-200 deficient cell lines, miR-200 precursors are detected in nuclear but not cytoplasmic compartment of these cells, indicating that promoter hypermethylation is not sufficient to explain the deficit of mature miR-200s. The ribonuclease Monocyte Chemotactic Protein-induced Protein-1 (MCPiP1) may counteract Dicer1 in miRNA maturation process. MCPiP1/Dicer1 mRNA and protein ratios appear higher in miR-200 deficient compared to miR-200 proficient cells, suggesting that MCPiP1 may compete with Dicer1 in mature miR-200 deficient cells. Inhibition of MCPiP1 allows the detection of miR-200 precursors in cytoplasm of miR-200 deficient cells, confirming its involvement in the loss of miR-200s. Also, reversion of MCPiP1/Dicer1 ratio by over-expression of Dicer1 in miR-200 deficient cells leads to the recovery of mature miR-200s. Finally, whereas human malignant pancreatic tissues (PDACs) express lower miR-200 levels than non malignant tissues (non-MPDs), MCPiP1/Dicer1 ratio appears higher in PDACs, when compared to non-MPDs, supporting the hypothesis that MCPiP1/Dicer1 ratio is determinant in regulating miR-200 maturation process in a subset of tumoral pancreatic cells., Competing Interests: CONFLICTS OF INTEREST The authors declare no conflicts of interest.
- Published
- 2018
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13. MMP2 and MMP9 as candidate biomarkers to monitor bevacizumab therapy in high-grade glioma.
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Tabouret E, Boudouresque F, Farina P, Barrié M, Bequet C, Sanson M, and Chinot O
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- Adult, Aged, Bevacizumab blood, Biomarkers, Tumor blood, Brain Neoplasms blood, Brain Neoplasms diagnosis, Disease Progression, Female, Glioblastoma blood, Glioblastoma diagnosis, Humans, Male, Middle Aged, Retrospective Studies, Treatment Outcome, Angiogenesis Inhibitors therapeutic use, Bevacizumab therapeutic use, Brain Neoplasms drug therapy, Glioblastoma drug therapy, Matrix Metalloproteinase 2 blood, Matrix Metalloproteinase 9 blood
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- 2015
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14. Differential expression of miR200a-3p and miR21 in grade II-III and grade IV gliomas: evidence that miR200a-3p is regulated by O⁶-methylguanine methyltransferase and promotes temozolomide responsiveness.
- Author
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Berthois Y, Delfino C, Metellus P, Fina F, Nanni-Metellus I, Al Aswy H, Pirisi V, Ouafik L, and Boudouresque F
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- Adult, Aged, Aged, 80 and over, Brain Neoplasms metabolism, Cell Line, Tumor, Cell Proliferation drug effects, Dacarbazine pharmacology, Female, Glioblastoma metabolism, Humans, Male, MicroRNAs genetics, Middle Aged, Neoplasm Grading, Temozolomide, Transcriptome, Antineoplastic Agents, Alkylating pharmacology, Brain Neoplasms pathology, DNA Modification Methylases metabolism, DNA Repair Enzymes metabolism, Dacarbazine analogs & derivatives, Drug Resistance, Neoplasm, Glioblastoma pathology, MicroRNAs metabolism, Tumor Suppressor Proteins metabolism
- Abstract
Glioblastoma multiforme (GBM) is the most common primary brain tumor and is among the deadliest of human cancers. Dysregulation of microRNAs (miRNAs) expression is an important step in tumor progression as miRNAs can act as tumor suppressors or oncogenes and may affect cell sensitivity to chemotherapy. Whereas the oncogenic miR21 has been shown to be overexpressed in gliomas, the expression and function of the tumor-supressor miR200a in GBMs remains unknown. In this study, we show that miR21 is upregulated in grade IV (GBMs) vs. grade II-III (LGs) gliomas, confirming that miR21 expression level is correlated with tumor grade, and that it may be considered as a marker of tumor progression. Conversely, miR200a is demonstrated for the first time to be downregulated in GBMs compared with LGs, and overexpression of miR200a in GBM cells is shown to promote TMZ-sensitivity. Interestingly, miR200a but not miR21 expression level is significantly higher in TMZ-responsive vs. -unresponsive tumoral glial cells in primary culture. Furthermore, miR200a appears negatively correlated with the expression of the DNA repair enzyme O (6)-methylguanine methyltransferase (MGMT), and the inhibition of MGMT activity results in an increase of miR200a expression in GBM cells. Taken together, these data strongly suggest that miR200a is likely to act as a crucial antitumoral factor regarding glioma progression. Interplay between miR200a and MGMT should be considered as potential mechanism involved in therapeutic response.
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- 2014
- Full Text
- View/download PDF
15. Association of matrix metalloproteinase 2 plasma level with response and survival in patients treated with bevacizumab for recurrent high-grade glioma.
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Tabouret E, Boudouresque F, Barrie M, Matta M, Boucard C, Loundou A, Carpentier A, Sanson M, Metellus P, Figarella-Branger D, Ouafik L, and Chinot O
- Subjects
- Adult, Aged, Bevacizumab, Biomarkers, Brain Neoplasms blood, Female, Glioblastoma blood, Humans, Male, Middle Aged, Neoplasm Recurrence, Local drug therapy, Survival Analysis, Treatment Outcome, Young Adult, Angiogenesis Inhibitors therapeutic use, Antibodies, Monoclonal, Humanized therapeutic use, Brain Neoplasms drug therapy, Brain Neoplasms mortality, Glioblastoma drug therapy, Glioblastoma mortality, Matrix Metalloproteinase 2 blood
- Abstract
Background: A predictive marker of bevacizumab activity is an unmet medical need. We evaluated the predictive value of selected circulating prebiomarkers involved in neoangiogenesis and invasion on patient outcome in recurrent high-grade glioma treated with bevacizumab., Methods: Analyzed in plasma were a set of 11 prebiomakers of interest (vascular endothelial growth factor receptor [VEGF]; VEGF receptor 2; basic fibroblast growth factor; stromal cell derived factor 1; placenta growth factor; urokinase-type plasminogen activator; plasminogen activator inhibitor 1; matrix metalloproteinases 2, 7, and 9; and adrenomedulline), using ELISA, at baseline and 2 weeks after bevacizumab initiation in a prospective cohort of 26 patients (Cohort 1). Correlations were validated in a separate retrospective cohort (Cohort 2; n = 50) and tested in cohort patients treated with cytotoxic agents without bevacizumab (Cohort 3; n = 34). Dosages were correlated to objective response, progression-free survival (PFS), and overall survival (OS)., Results: In Cohort 1, high MMP2 baseline level was associated with a probability of objective response of 83.3% versus 15.4% for low MMP2 level (P = .001). In multivariate analysis, baseline level of MMP2 correlated with PFS (hazard ratio, 3.92; 95% confidence interval [CI]:1.46-10.52; P = .007) and OS (hazard ratio, 4.62; 95% CI: 1.58-13.53; P = .005), as decrease of VEGF (P = .038 for PFS and P = .013 for OS) and MMP9 (P = .016 for PFS and P = .025 for OS). In Cohort 2, MMP2, but not MMP9, confirmed its predictive significance. In Cohort 3, no association was found between MMP2, MMP9, and outcome., Conclusion: In patients with recurrent high-grade glioma treated with bevacizumab, but not with cytotoxic agent, high MMP2 plasma levels are associated with prolonged tumor control and survival. MMP2 should be tested in randomized clinical trials that evaluate bevacizumab efficacy, and its biological role reassessed.
- Published
- 2014
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16. Adrenomedullin blockade suppresses growth of human hormone-independent prostate tumor xenograft in mice.
- Author
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Berenguer-Daizé C, Boudouresque F, Bastide C, Tounsi A, Benyahia Z, Acunzo J, Dussault N, Delfino C, Baeza N, Daniel L, Cayol M, Rossi D, El Battari A, Bertin D, Mabrouk K, Martin PM, and Ouafik L
- Subjects
- Adrenomedullin antagonists & inhibitors, Adrenomedullin immunology, Androgens, Animals, Antibodies immunology, Apoptosis immunology, Calcitonin Receptor-Like Protein, Carrier Proteins metabolism, Castration, Cell Line, Tumor, Cell Movement immunology, Cell Proliferation, Cyclic AMP metabolism, Endothelial Cells immunology, Humans, Male, Mice, Mice, Nude, Neoplasm Invasiveness, Neoplasm Transplantation, Pericytes immunology, Receptor Activity-Modifying Protein 2 metabolism, Receptor Activity-Modifying Protein 3 metabolism, Receptors, Adrenomedullin immunology, Receptors, Calcitonin metabolism, Signal Transduction immunology, Transplantation, Heterologous, Adrenomedullin metabolism, Lymphangiogenesis, Neovascularization, Pathologic, Prostatic Neoplasms, Castration-Resistant metabolism
- Abstract
Purpose: To study the role of the adrenomedullin system [adrenomedullin and its receptors (AMR), CLR, RAMP2, and RAMP3] in prostate cancer androgen-independent growth., Experimental Design: Androgen-dependent and -independent prostate cancer models were used to investigate the role and mechanisms of adrenomedullin in prostate cancer hormone-independent growth and tumor-associated angiogenesis and lymphangiogenesis., Results: Adrenomedullin and AMR were immunohistochemically localized in the carcinomatous epithelial compartment of prostate cancer specimens of high grade (Gleason score >7), suggesting a role of the adrenomedullin system in prostate cancer growth. We used the androgen-independent Du145 cells, for which we demonstrate that adrenomedullin stimulated cell proliferation in vitro through the cAMP/CRAF/MEK/ERK pathway. The proliferation of Du145 and PC3 cells is decreased by anti-adrenomedullin antibody (αAM), supporting the fact that adrenomedullin may function as a potent autocrine/paracrine growth factor for prostate cancer androgen-independent cells. In vivo, αAM therapy inhibits the growth of Du145 androgen-independent xenografts and interestingly of LNCaP androgen-dependent xenografts only in castrated animals, suggesting strongly that adrenomedullin might play an important role in tumor regrowth following androgen ablation. Histologic examination of αAM-treated tumors showed evidence of disruption of tumor vascularity, with depletion of vascular as well as lymphatic endothelial cells and pericytes, and increased lymphatic endothelial cell apoptosis. Importantly, αAM potently blocks tumor-associated lymphangiogenesis, but does not affect established vasculature and lymphatic vessels in normal adult mice., Conclusions: We conclude that expression of adrenomedullin upon androgen ablation in prostate cancer plays an important role in hormone-independent tumor growth and in neovascularization by supplying/amplifying signals essential for pathologic neoangiogenesis and lymphangiogenesis. Clin Cancer Res; 19(22); 6138-50. ©2013 AACR.
- Published
- 2013
- Full Text
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17. Neutralization of adrenomedullin inhibits the growth of human glioblastoma cell lines in vitro and suppresses tumor xenograft growth in vivo.
- Author
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Ouafik L, Sauze S, Boudouresque F, Chinot O, Delfino C, Fina F, Vuaroqueaux V, Dussert C, Palmari J, Dufour H, Grisoli F, Casellas P, Brünner N, and Martin PM
- Subjects
- Adrenomedullin, Animals, Antibodies pharmacology, Cell Division drug effects, Cell Division physiology, Glioblastoma metabolism, Glioma metabolism, Glioma pathology, Humans, Immunohistochemistry, Male, Mice, Mice, Inbred Strains, Mixed Function Oxygenases genetics, Multienzyme Complexes genetics, Peptide Fragments pharmacology, Peptides genetics, Peptides immunology, Peptides metabolism, RNA, Messenger metabolism, Tumor Cells, Cultured, Glioblastoma pathology, Neoplasm Transplantation, Peptides physiology, Transplantation, Heterologous
- Abstract
Presently, there is no effective treatment for glioblastoma, the most malignant and common brain tumor. Growth factors are potential targets for therapeutic strategies because they are essential for tumor growth and progression. Peptidylglycine alpha-amidating monooxygenase is the enzyme producing alpha-amidated bioactive peptides from their inactive glycine-extended precursors. The high expression of peptidylglycine alpha-amidating monooxygenase mRNA in glioblastoma and glioma cell lines points to the involvement of alpha-amidated peptides in tumorigenic growth processes in the brain. After screening of amidated peptides, it was found that human glioblastoma cell lines express high levels of adrenomedullin (AM) mRNA, and that immunoreactive AM is released into the culture medium. AM is a multifunctional regulatory peptide with mitogenic and angiogenic capabilities among others. Real-time quantitative reverse transcriptase-polymerase chain reaction analysis showed that AM mRNA was correlated to the tumor type and grade, with high expression in all glioblastomas analyzed, whereas a low expression was found in anaplastic astrocytomas and barely detectable levels in low-grade astrocytomas and oligodendrogliomas. In the present study we also demonstrate the presence of mRNA encoding the putative AM receptors, calcitonin receptor-like receptor/receptor activity-modifying protein-2 and -3 (CRLR/RAMP2; CRLR/RAMP3) in both glioma tissues and glioblastoma cell lines and further show that exogenously added AM can stimulate the growth of these glioblastoma cells in vitro. These findings suggest that AM may function as an autocrine growth factor for glioblastoma cells. One way to test the autocrine hypothesis is to interrupt the function of the endogenously produced AM. Herein, we demonstrate that a polyclonal antibody specific to AM, blocks the binding of the hormone to its cellular receptors and decreases by 33% (P < 0.001) the growth of U87 glioblastoma cells in vitro. Intratumoral administration of the anti-AM antibody resulted in a 70% (P < 0.001) reduction in subcutaneous U87 xenograft weight 21 days after treatment. Furthermore, the density of vessels was decreased in the antibody-treated tumors. These findings support that AM may function as a potent autocrine/paracrine growth factor for human glioblastomas and demonstrate that inhibition of the action of AM (produced by tumor cells) may suppress tumor growth in vivo.
- Published
- 2002
- Full Text
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18. Expression of adrenomedullin and peptide amidation activity in human prostate cancer and in human prostate cancer cell lines.
- Author
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Rocchi P, Boudouresque F, Zamora AJ, Muracciole X, Lechevallier E, Martin PM, and Ouafik L
- Subjects
- Adenocarcinoma enzymology, Adrenomedullin, Animals, Cell Division drug effects, Humans, Immunohistochemistry, In Situ Hybridization, Male, Mice, Mice, Nude, Mixed Function Oxygenases biosynthesis, Mixed Function Oxygenases genetics, Neoplasm Proteins metabolism, Neoplasm Transplantation, Neoplasms, Hormone-Dependent enzymology, Neoplasms, Hormone-Dependent metabolism, Peptides genetics, Peptides pharmacology, Phosphopyruvate Hydratase metabolism, Prostatic Hyperplasia enzymology, Prostatic Hyperplasia metabolism, Prostatic Neoplasms enzymology, RNA, Messenger biosynthesis, RNA, Messenger genetics, Reverse Transcriptase Polymerase Chain Reaction, Transplantation, Heterologous, Tumor Cells, Cultured, Adenocarcinoma metabolism, Mixed Function Oxygenases metabolism, Multienzyme Complexes, Peptides metabolism, Prostatic Neoplasms metabolism
- Abstract
After therapeutic hormone deprivation, prostate cancer (CaP) cells often develop androgen-independent growth through not-well-defined mechanisms. The presence of neuroendocrine (NE) cells is often greater in prostate carcinoma than in normal prostate, and the frequency of NE cells correlates with tumor malignancy, loss of androgen sensitivity, increase of autocrine-paracrine activity, and poor prognosis. In some CaPs, neuropeptides have been previously implicated as growth factors. Peptidylglycine alpha-amidating monooxygenase (PAM) is the enzyme producing alpha-amidated bioactive peptides from their inactive glycine-extended precursors. In the present work, we demonstrate that androgen-independent PC-3 and DU145 cell lines, derived from human CaP, express PAM in vitro and in xenografts implanted in athymic nude mice, indicating that they are able to produce alpha-amidated peptides. Contrarily, barely detectable levels of PAM were found in the androgen-sensitive LNCaP cell line. We also show that whereas PC-3 and DU145 cells produce and secrete adrenomedullin (AM), a multifunctional amidated peptide, no expression was found in LNCaP cells. We further demonstrate that AM acts as a growth factor for DU145 cells, which suggests the existence of an autocrine loop mechanism that could potentially drive neoplastic growth. PAM mRNA levels were found to be 3-fold higher in prostate adenocarcinomas compared with that of human benign prostate hyperplasia (BPH) as demonstrated by real-time quantitative reverse transcription-PCR. The analysis of AM message expression in BPH and CaP (Gleason's score, 6-9) shows a clear distinction between benign and CaP. The expression was detected only in adenocarcinomas tissues with a marked increase in samples with a high Gleason's score. Immunocytochemically, AM was localized in the carcinomatous epithelial compartment. NE phenotype, assessed after the immunocytochemical localization of neuron-specific enolase (NSE), was found in both the epithelial and the stromal compartments of cancers; in BPH, only some spare basal cells were NSE-labeled. Cancer progression could be accelerated by peptides secreted by a population of cells capable of inducing androgen-independent tumoral growth via autocrine-paracrine mechanisms.
- Published
- 2001
19. Evidence of high expression of peptidylglycine alpha-amidating monooxygenase in the rat uterus: estrogen regulation.
- Author
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El Meskini R, Delfino C, Boudouresque F, Oliver C, Martin PM, and Ouafik L'H
- Subjects
- Animals, Female, In Situ Hybridization, RNA, Messenger analysis, Rats, Rats, Sprague-Dawley, Estrogens physiology, Mixed Function Oxygenases physiology, Multienzyme Complexes, Receptors, Estrogen physiology, Uterus physiology
- Abstract
In the present study, high levels of peptidylglycine alpha-amidating monooxygenase (PAM), which catalyzes the two-step formation of bioactive alpha-amidated peptides from their glycine-extended precursors, have been found in the uterus. Expression of PAM was evaluated in the uterus of intact cycling adult female rats and after experimental manipulation of the estrogen status of the rats. During the estrous cycle, PAM mRNA levels exhibited striking changes inversely related to the physiological variations of plasma estrogen levels. The levels of PAM transcripts changed markedly during the estrous cycle, reaching the highest levels at metestrus. There was a 15-fold increase in the abundance of PAM mRNA between metestrus and proestrus. Chronic treatment of ovariectomized rats with 17beta-estradiol decreased PAM mRNA levels to values comparable with those found in intact rats at proestrus. Progesterone was without effect on PAM mRNA levels, indicating that the effect was specific for estradiol. In situ hybridization studies were conducted to determine the tissue disposition and cell types expressing PAM. High levels of PAM mRNA were localized in the endometrium at the level of luminal and glandular cells. A weak signal was observed in stromal cells, and the myometrium cells were negative. 17beta-Estradiol treatment induced an overall decrease of the hybridization signal, as compared with ovariectomized rats. These results demonstrate the presence of high levels of PAM in the uterus and indicate that estrogens are involved in regulating the expression of the enzyme in this tissue. However, the present study provides no information regarding whether this regulation takes place at the level of transcription or influences mRNA stability.
- Published
- 1998
- Full Text
- View/download PDF
20. Identification of a novel cis-element in the 3'-untranslated region of mammalian peptidylglycine alpha-amidating monooxygenase messenger ribonucleic acid.
- Author
-
Fraboulet S, Boudouresque F, Delfino C, and Ouafik L
- Subjects
- Animals, Base Sequence, Binding Sites, Conserved Sequence, Molecular Sequence Data, Rats, Mixed Function Oxygenases genetics, Multienzyme Complexes, RNA, Messenger metabolism, RNA-Binding Proteins analysis
- Abstract
Peptidylglycine alpha-amidating monooxygenase (PAM; EC 1.14.17.3) catalyzes the COOH-terminal alpha-amidation of peptidylglycine substrates, yielding amidated products. Growing evidence suggests that the metabolism of PAM messenger RNAs (mRNAs) can be regulated within the cytoplasm. To understand the mechanisms controlling the metabolism of PAM mRNAs, we sought to identify cis elements of the 3'-untranslated region (3'-UTR) of PAM mRNA that are recognized by cytoplasmic factors. From gel retardation assays, one sequence element is shown to form a specific RNA-protein complex. The protein-binding site of the complex was determined by ribonuclease T1 mapping, by blocking the putative binding site with antisense oligonucleotide, and by competition assays. Using 3'-end-labeled RNA in gel shift and UV cross-linking analyses, we detected in the 3'-UTR a novel 20-nucleotide cis element that interacted with a widely distributed cellular cytosolic protease-sensitive factor(s) to form a 60-kDa PAM mRNA-binding protein complex. The binding activity was redox sensitive. Tissue distribution of the protein in the rat showed a marked tissue-specific expression, with ovary, testis, lung, heart septum, anterior pituitary and hypothalamus containing large amounts compared with liver, ventricle, atrium, and neurointermediate lobe. No binding activity was detectable in pancreas, intestine, or kidney extracts. Northwestern blot analysis of AtT-20 (mouse corticotrope tumor cell line) cytoplasmic extracts revealed a protein of 46 kDa. Thus, we have identified a widely distributed cellular protein that binds to a conserved domain within the 3'-UTR of PAM mRNA from many animal species. Although these data suggest that cis element-binding activity could be a cytoplasmic regulator of PAM mRNA metabolism, the functional consequences of this binding remain to be determined.
- Published
- 1998
- Full Text
- View/download PDF
21. Estrogen regulation of peptidylglycine alpha-amidating monooxygenase messenger ribonucleic acid levels by a nuclear posttranscriptional event.
- Author
-
El Meskini R, Boudouresque F, and Ouafik L
- Subjects
- Animals, Culture Techniques, Cytoplasm metabolism, Drug Stability, Estradiol pharmacology, Female, Half-Life, Ovariectomy, Pituitary Gland, Anterior metabolism, Poly A metabolism, RNA Precursors metabolism, RNA, Messenger antagonists & inhibitors, RNA, Messenger chemistry, Rats, Rats, Sprague-Dawley, Transcription, Genetic drug effects, Cell Nucleus metabolism, Estrogens physiology, Mixed Function Oxygenases genetics, Multienzyme Complexes, Protein Processing, Post-Translational physiology, RNA, Messenger metabolism
- Abstract
Peptidylglycine alpha-amidating monooxygenase (PAM; EC 1.14.17.3) is a bifunctional protein containing two enzymes that act sequentially to catalyze the conversion of glycine-extended peptides into COOH-terminal amidated peptides. We have previously shown that PAM messenger RNA (mRNA) levels in the anterior pituitary of intact cycling adult female rats showed changes inversely related to the physiological variations of plasma estrogen levels during the estrous cycle. Chronic treatment of ovariectomized (OVX) rats with 17beta-estradiol was accompanied by a 4.5 +/- 0.5-fold decrease in total PAM mRNA and a 2-fold decrease in PAM activity in the anterior pituitary gland. To investigate the cellular site at which 17beta-estradiol acts to affect the PAM mRNA, we made parallel measurements of the relative levels of PAM mRNA and nuclear precursor RNA and the relative rate of gene transcription after treatments designed to alter the estrogen status. The transcription rate experiments indicated that these 17beta-estradiol effects were not due to reduced PAM gene activity, suggesting that a posttranscriptional mechanism was involved. The most common mechanism of posttranscriptional regulation affects cytoplasmic mRNA stability. Primary rat pituitary cell cultures from OVX and OVX-17beta-estradiol-treated rats in the presence of actinomycin D showed that 17beta-estradiol treatment decreased the half-life of PAM mRNA from 15-16 h to 8-9 h. There was no effect of 17beta-estradiol on PAM mRNA poly(A) tail length or site of polyadenylation. However, in this study the down-regulation of PAM was identified as a nuclear event. Analysis of nuclear RNA with probes specific for PAM intron sequences shows that decreased PAM expression after 17beta-estradiol treatment was largely due to intranuclear destabilization of the primary transcript. The levels of nuclear precursor RNA were decreased roughly 5- to 6-fold in OVX + 17beta-estradiol compared with OVX rats. The decrease in PAM mRNA is blocked by cycloheximide, indicating that its requires new protein synthesis. Mechanisms that would generate such an effect include altered stability of unprocessed message in the nucleus. The proportional changes observed in the nuclear precursor and mRNA levels suggest that the site of control is at the level of stability of the nuclear precursor RNA for PAM mRNA.
- Published
- 1997
- Full Text
- View/download PDF
22. Estrogen regulation of peptidylglycine alpha-amidating monooxygenase expression in anterior pituitary gland.
- Author
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el Meskini R, Delfino C, Boudouresque F, Hery M, Oliver C, and Ouafik L
- Subjects
- Alternative Splicing, Animals, Estrus metabolism, Female, In Situ Hybridization, Ovariectomy, Progesterone pharmacology, Rats, Rats, Sprague-Dawley, Estradiol pharmacology, Gene Expression Regulation, Enzymologic drug effects, Mixed Function Oxygenases genetics, Multienzyme Complexes, Pituitary Gland, Anterior enzymology
- Abstract
The pituitary is a rich source of peptidylglycine alpha-amidating monooxygenase (PAM). This bifunctional protein contains peptidylglycine alpha-hydroxylating monooxygenase (PHM) and peptidyl-alpha-hydroxyglycine alpha-amidating lyase catalytic domains necessary for the two-step formation of alpha-amidated peptides from their COOH-terminal glycine extended precursors. Expression of PAM was evaluated in the anterior pituitary of intact cycling adult female rat and after experimental manipulation of estrogen status. PAM messenger RNA (mRNA) levels showed changes inversely related to the physiological variations of plasma estrogen levels during the estrous cycle. Chronic treatment of ovariectomized (OVX) rats with 17 beta-estradiol decreased PAM mRNA levels to values comparable with those found in intact rats at proestrus. In situ hybridization of anterior pituitary sections using 35S-labeled full length RNA antisense transcripts of rat PAM-1 complementary DNA showed that 17 beta-estradiol treatment induced an overall decrease of the hybridization signal, as compared with OVX rats. Progesterone treatment did not change PAM mRNA levels both in OVX or OVX + E2 rats. Based on Northern blot analysis and amplification of fragments derived from rat PAM-1 by RT-PCR, it was found that estrogen status does not affect the distribution of PAM mRNA among its various alternatively spliced forms. In OVX 17 beta-estradiol treated rats, the specific activity of PAM in the anterior pituitary decreased in both soluble and particulate fractions compared with OVX animals. Western blot analysis demonstrated a 105-kDa PAM protein in particulate fractions prepared from OVX and OVX-17 beta-estradiol treated animals. The soluble fraction from OVX animals contained major PAM proteins of 105, 95, 84, 75, and 45 kDa, and 17 beta-estradiol treatment caused a decrease in the prevalence of these proteins. These results indicate that estrogens are involved, either directly or indirectly, in regulating the expression of PAM in several cell types in the anterior pituitary gland.
- Published
- 1997
- Full Text
- View/download PDF
23. Insulin-induced hypoglycemia stimulates corticotropin-releasing factor and arginine vasopressin secretion into hypophysial portal blood of conscious, unrestrained rams.
- Author
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Caraty A, Grino M, Locatelli A, Guillaume V, Boudouresque F, Conte-Devolx B, and Oliver C
- Subjects
- Adrenocorticotropic Hormone blood, Animals, Blood Glucose metabolism, Hydrocortisone blood, Male, Pituitary Gland blood supply, Secretory Rate drug effects, Sheep, Arginine Vasopressin metabolism, Corticotropin-Releasing Hormone metabolism, Hypoglycemia physiopathology, Insulin pharmacology
- Abstract
Insulin-induced hypoglycemia (IIH) is a strong stimulator of pituitary ACTH secretion. The mechanisms by which IIH activates the corticotrophs are still controversial. Indeed, in rats the variations of corticotropin-releasing factor (CRF) and arginine vasopressin (AVP) secretion in hypophysial portal blood (HPB) during IIH have been diversely appreciated. This may be due to the stressful conditions required for portal blood collection in rats. We studied the effects of IIH on the secretion of CRF and AVP in HPB and on the release of ACTH and cortisol in peripheral plasma in conscious, unrestrained, castrated rams. After the injection of a low (0.2 IU/kg) or high dose (2 IU/kg) of insulin, ACTH and cortisol levels in peripheral plasma increased in a dose-related manner. After injection of the low dose of insulin, CRF and AVP secretion in HPB were equally stimulated. After injection of the high dose of insulin, CRF secretion was further stimulated, while AVP release was dramatically increased. These results suggest that when the hypoglycemia is moderate, CRF is the main factor triggering ACTH release, and that the increased AVP secretion potentiates the stimulatory effect of CRF. When hypoglycemia is deeper, AVP secretion becomes predominant and may by itself stimulate ACTH release.
- Published
- 1990
- Full Text
- View/download PDF
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