RESUMO
The hypothesis that proximity to the Sun causes variation of decay constants at permille level has been tested and disproved. Repeated activity measurements of mono-radionuclide sources were performed over periods from 200 days up to four decades at 14 laboratories across the globe. Residuals from the exponential nuclear decay curves were inspected for annual oscillations. Systematic deviations from a purely exponential decay curve differ from one data set to another and are attributable to instabilities in the instrumentation and measurement conditions. The most stable activity measurements of alpha, beta-minus, electron capture, and beta-plus decaying sources set an upper limit of 0.0006% to 0.008% to the amplitude of annual oscillations in the decay rate. Oscillations in phase with Earth's orbital distance to the Sun could not be observed within a 10-6 to 10-5 range of precision. There are also no apparent modulations over periods of weeks or months. Consequently, there is no indication of a natural impediment against sub-permille accuracy in half-life determinations, renormalisation of activity to a distant reference date, application of nuclear dating for archaeology, geo- and cosmochronology, nor in establishing the SI unit becquerel and seeking international equivalence of activity standards.
RESUMO
BACKGROUND: Lenalidomide is an active immunomodulatory and antiproliferative agent in multiple myeloma. However, the molecular mechanisms driving these activities are not yet fully elucidated. Therefore, we investigated the modulation of the cytokine/growth factor patterns of myeloma cells under LEN treatment. METHODS: Lenalidomide effect on myeloma cell proliferation was investigated in a myeloma cell line collection (n=23) by (3)H-thymidine incorporation. Modulation of the cytokine/growth factor patterns of myeloma cells under LEN treatment was analysed by real-time quantitative PCR. RESULTS: Lenalidomide inhibits the proliferation of two-thirds of myeloma cell lines independently of their genetic background. We demonstrated that LEN increased TNF-α and IL-8 inflammatory cytokines and insulin-like growth factor-1 (IGF-1) growth factor in both sensitive and resistant myeloma cells to LEN. CONCLUSION: Lenalidomide favours a uniform TNF-α and IL-8 inflammatory and IGF-1 secretory profile of myeloma cells, an observation that raises important questions for therapeutic approaches incorporating the agent.
Assuntos
Inibidores da Angiogênese/farmacologia , Antineoplásicos/farmacologia , Fator de Crescimento Insulin-Like I/metabolismo , Interleucina-8/metabolismo , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/metabolismo , Talidomida/análogos & derivados , Fator de Necrose Tumoral alfa/metabolismo , Células da Medula Óssea/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos , Humanos , Fatores Imunológicos/farmacologia , Lenalidomida , Talidomida/farmacologiaRESUMO
BACKGROUND: Cancer cells are frequently addicted to deregulated oncogenic protein translation. The small molecule 4EG-I selectively inhibits the cap-dependent translation of mRNAs. As multiple myeloma is an incurable disease that requires new therapeutic approaches, we investigated whether targeting the translation initiation pathway could be a target for myeloma therapy. METHODS: Six myeloma cell lines and primary samples were included in this study. The 4EGI-1 effect was determined by AnnexinV staining and caspase activation. Modification of Bcl-2 protein expression was analysed, and the significance of modified proteins was analysed by knock-down experiments. RESULTS: We demonstrated that 4EGI-1 impaired the assembly of the eIF4F complex and decreased the expression of the eIF4E-regulated proteins in myeloma cells. Furthermore, we showed that 4EGI-1 induced strong apoptosis in five out of six myeloma cell lines. Apoptosis is associated with the activation of the intrinsic mitochondrial pathway. The 4EGI-1 triggered Noxa induction only in cells undergoing apoptosis through endoplasmic reticulum (ER) stress. Furthermore, Noxa silencing prevented myeloma cells from 4EGI-1-induced apoptosis. Finally, Noxa induction led to a disruption of Mcl-1/Bim complexes in parallel to the generation of 'Mcl-1-free Noxa'. CONCLUSION: Our results suggested that the use of inhibitors that directly target the translation initiation complex eIF4F could represent a potential novel approach for multiple myeloma therapy.
Assuntos
Apoptose/efeitos dos fármacos , Fator de Iniciação 4F em Eucariotos/antagonistas & inibidores , Mieloma Múltiplo/tratamento farmacológico , Nitrocompostos/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/fisiologia , Tiazóis/farmacologia , Proteínas Reguladoras de Apoptose/metabolismo , Proteína 11 Semelhante a Bcl-2 , Linhagem Celular Tumoral , Fator de Iniciação 4F em Eucariotos/fisiologia , Humanos , Hidrazonas , Proteínas de Membrana/metabolismo , Mieloma Múltiplo/patologia , Proteína de Sequência 1 de Leucemia de Células Mieloides , Iniciação Traducional da Cadeia Peptídica/efeitos dos fármacos , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/análise , Proteínas Proto-Oncogênicas c-bcl-2/metabolismoRESUMO
BACKGROUND: although gene expression profile of multiple myeloma (MM) patients shows a wide range of Bik/Nbk expression, varying from absent to high, its regulation and function in myeloma cells is poorly understood. Thus, we addressed these questions in MM. METHODS: human myeloma cell lines (HMCLs) and primary purified myeloma cells were studied for Bcl-2 family protein expression by western blot and further correlation analysis was performed. Correlative study between Bik and thyrotroph embryonic factor (TEF) transcription factor expression was analysed by PCR. Stress oxidative response was analysed by flow cytometry. RESULTS: a strong expression of Bik protein was found only in one out of three of HMCL and correlated to Bcl-2 expression (P=0.0006). We demonstrated that Bik could be regulated at the protein level by Bcl-2 and at the transcriptional level by TEF. Bik overexpression sensitises myeloma cells to oxidative stress whereas Bik silencing increases resistance to H(2)O(2) oxidative stress. Furthermore, Bik ectopic expression disrupts Bim/Bcl-2 and Bim/Bcl-xL endogenous complexes triggering Bim release that could induce Bax and Bak activation. CONCLUSIONS: ours results suggest that Bik has a role in both, apoptosis induction and sensitivity to oxidative stress in myeloma cells. Small BH3 mimetic molecules should be considered for further apoptosis-based therapy in myeloma cells expressing endogenous Bik/Bcl-2 complexes.
Assuntos
Proteínas Reguladoras de Apoptose/fisiologia , Apoptose , Proteínas de Membrana/fisiologia , Mieloma Múltiplo/patologia , Estresse Oxidativo , Proteínas Reguladoras de Apoptose/genética , Fatores de Transcrição de Zíper de Leucina Básica/genética , Humanos , Proteínas de Membrana/genética , Proteínas Mitocondriais , Mieloma Múltiplo/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/análise , RNA Mensageiro/análise , Proteína Supressora de Tumor p53/análiseRESUMO
The humanised form of an antagonistic anti-IGF-1R mAb (AVE1642) selectively inhibits the growth of CD45(neg) myeloma cells. AVE1642 strongly increased bortezomib-induced apoptosis, correlated with an increase of Noxa expression. These results support the therapeutic use of anti-IGF-1R/bortezomib in CD45(neg) Myeloma patients, particularly those with the most aggressive form, t(4,14).
Assuntos
Anticorpos Monoclonais/farmacologia , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Ácidos Borônicos/farmacologia , Antígenos Comuns de Leucócito/metabolismo , Mieloma Múltiplo/patologia , Pirazinas/farmacologia , Receptor IGF Tipo 1/imunologia , Apoptose/imunologia , Bortezomib , Caspase 3/metabolismo , Proliferação de Células/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Humanos , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/imunologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Células Tumorais CultivadasRESUMO
Selective inhibition of BCL-2 is expected to enhance therapeutic vulnerability in luminal estrogen receptor-positive breast cancers. We show here that the BCL-2 dependency of luminal tumor cells is nevertheless mitigated by breast cancer-associated fibroblasts (bCAFs) in a manner that defines MCL-1 as another critical therapeutic target. bCAFs favor MCL-1 expression and apoptotic resistance in luminal cancer cells in a IL-6 dependent manner while their own, robust, survival also relies on MCL-1. Studies based on ex vivo cultures of human luminal breast cancer tissues further argue that the contribution of stroma-derived signals to MCL-1 expression shapes BCL-2 dependency. Thus, MCL-1 inhibitors are beneficial for targeted apoptosis of breast tumor ecosystems, even in a subtype where MCL-1 dependency is not intrinsically driven by oncogenic pathways.
Assuntos
Neoplasias da Mama/metabolismo , Fibroblastos Associados a Câncer/metabolismo , Proteína de Sequência 1 de Leucemia de Células Mieloides/deficiência , Proteína de Sequência 1 de Leucemia de Células Mieloides/metabolismo , Receptores de Estrogênio/metabolismo , Apoptose/fisiologia , Neoplasias da Mama/patologia , Fibroblastos Associados a Câncer/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/fisiologia , Feminino , Humanos , Interleucina-6/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismoRESUMO
Mitochondria are involved in hematopoietic cell homeostasis through multiple ways such as oxidative phosphorylation, various metabolic processes and the release of cytochrome c in the cytosol to trigger caspase activation and cell death. In erythroid cells, the mitochondrial steps in heme synthesis, iron (Fe) metabolism and Fe-sulfur (Fe-S) cluster biogenesis are of particular importance. Mutations in the specific delta-aminolevulinic acid synthase (ALAS) 2 isoform that catalyses the first and rate-limiting step in heme synthesis pathway in the mitochondrial matrix, lead to ineffective erythropoiesis that characterizes X-linked sideroblastic anemia (XLSA), the most common inherited sideroblastic anemia. Mutations in the adenosine triphosphate-binding cassette protein ABCB7, identified in XLSA with ataxia (XLSA-A), disrupt the maturation of cytosolic (Fe-S) clusters, leading to mitochondrial Fe accumulation. In addition, large deletions in mitochondrial DNA, whose integrity depends on a specific DNA polymerase, are the hallmark of Pearson's syndrome, a rare congenital disorder with sideroblastic anemia. In acquired myelodysplastic syndromes at early stage, exacerbation of physiological pathways involving caspases and the mitochondria in erythroid differentiation leads to abnormal activation of a mitochondria-mediated apoptotic cell death pathway. In contrast, oncogenesis-associated changes at the mitochondrial level can alter the apoptotic response of transformed hematopoietic cells to chemotherapeutic agents. Recent findings in mitochondria metabolism and functions open new perspectives in treating hematopoietic cell diseases, for example various compounds currently developed to trigger tumor cell death by directly targeting the mitochondria could prove efficient as either cytotoxic drugs or chemosensitizing agents in treating hematological malignancies.
Assuntos
Doenças Hematológicas/fisiopatologia , Hematopoese/fisiologia , Mitocôndrias/fisiologia , Animais , Diferenciação Celular , Transformação Celular Neoplásica , Senescência Celular/genética , DNA Mitocondrial/genética , Doenças Hematológicas/patologia , Humanos , Ferro/metabolismo , Mitocôndrias/metabolismo , MutaçãoRESUMO
OBJECTIVE: (R,R,R)-alpha-tocopherol is a fat-soluble antioxidant vitamin generally ingested with other dietary antioxidants. The objective of this study was to assess whether the main dietary antioxidant classes, that is carotenoids, polyphenols, vitamin C and gamma-tocopherol, affect the intestinal absorption of alpha-tocopherol. METHODS, DESIGN AND SUBJECTS: We evaluated first the effect of different combinations of antioxidants on (R,R,R)-alpha-tocopherol absorption by a human intestinal cell line (Caco-2 clone TC7). Then we compared the effect of two doses of a dietary antioxidant (lutein) on the postprandial chylomicron alpha-tocopherol responses to an alpha-tocopherol-rich meal. Eight healthy men ate two similar meals in a random order at a 1 month interval. The meals contained 24 mg alpha-tocopherol in sunflower oil plus either 18 or 36 mg lutein. Blood samples were collected during the postprandial periods to compare chylomicron alpha-tocopherol responses. RESULTS: A mixture of polyphenols (gallic acid, caffeic acid, (+)-catechin and naringenin) and a mixture of carotenoids (lycopene, beta-carotene and lutein) significantly impaired alpha-tocopherol absorption in Caco-2 cells (P<0.001 and P<0.0001, respectively). The inhibitory effect of gamma-tocopherol was close to significance (P=0.055). In contrast, vitamin C had no significant effect (P=0.158). Naringenin was the only polyphenol that significantly impaired alpha-tocopherol absorption. Postprandial alpha-tocopherol response was weakest at the highest dose of lutein (616+/-280 nmol/l h vs 1001+/-287 nmol/l h). The observed extent of reduction (-38%, P=0.069) supported the inhibitory effect of carotenoids observed in the Caco-2 experiments. CONCLUSION: Naringenin, carotenoids and probably gamma-tocopherol can impair alpha-tocopherol absorption whereas vitamin C and phenolic acids have no effect.
Assuntos
Antioxidantes/farmacologia , Quilomícrons/química , Absorção Intestinal/efeitos dos fármacos , Luteína/farmacologia , alfa-Tocoferol/farmacocinética , Adulto , Área Sob a Curva , Ácido Ascórbico/farmacologia , Células CACO-2 , Carotenoides/farmacologia , Estudos Cross-Over , Relação Dose-Resposta a Droga , Flavonoides/farmacologia , Humanos , Masculino , Fenóis/farmacologia , Polifenóis , Período Pós-Prandial , gama-Tocoferol/farmacologiaRESUMO
TP53 deletion or mutation is frequent in B-cell malignancies and is associated with a low response rate. We describe here the p53 landscape in B-cell malignancies, from B-Acute Lymphoblastic Leukemia to Plasma Cell Leukemia, by analyzing incidence of gain or loss of function of actors both upstream and within the p53 pathway, namely MYC, RAS, ARF, MDM2, ATM and TP53. Abnormalities are not equally distributed and their incidence is highly variable among malignancies. Deletion and mutation, usually associated, of ATM or TP53 are frequent in Diffuse Large B-Cell Lymphoma and Mantle Cell Lymphoma. MYC gain, absent in post-GC malignancies, is frequent in B-Prolymphocytic-Leukemia, Multiple Myeloma and Plasma Cell Leukemias. RAS mutations are rare except in MM and PCL. Multiple Factorial Analysis notes that MYC deregulation is closely related to TP53 status. Moreover, MYC gain, TP53 deletion and RAS mutations are inversely correlated with survival. Based on this landscape, we further propose targeted therapeutic approaches for the different B-cell malignancies.
Assuntos
Regulação Neoplásica da Expressão Gênica , Linfoma de Células B/genética , Proteína Supressora de Tumor p53/genética , Animais , Linfócitos B/metabolismo , Linfócitos B/patologia , Humanos , Linfoma de Células B/metabolismo , Linfoma de Células B/patologia , Mutação , Transdução de Sinais , Proteína Supressora de Tumor p53/análise , Proteína Supressora de Tumor p53/metabolismoRESUMO
We and others have shown that Mcl-1 was essential for the survival of human myeloma cells in vitro. Furthermore, this antiapoptotic protein is upregulated by interleukin-6, which plays a critical role in multiple myeloma (MM). For these reasons, we have evaluated the expression of Mcl-1 in vivo in normal, reactive and malignant plasma cells (PC), that is, myeloma cells from 51 patients with MM and 21 human myeloma cell lines (HMCL) using flow cytometry. We show that Mcl-1 is overexpressed in MM in comparison with normal bone marrow PC. In total, 52% of patients with MM at diagnosis (P=0.017) and 81% at relapse (P=0.014 for comparison with diagnosis) overexpress Mcl-1. Of note, only HMCL but not reactive plasmacytoses have abnormal Mcl-1 expression, although both PC expansions share similar high proliferation rates. Of interest, Bcl-2 as opposed to Mcl-1, does not discriminate malignant from normal PC. Finally, the level of Mcl-1 expression is related to disease severity, the highest values at diagnosis being associated with the shortest event-free survival (P=0.002). In conclusion, Mcl-1, which has been shown to be essential for the survival of human myeloma cells in vitro, is overexpressed in vivo in MM in relation with relapse and shorter survival. Mcl-1 represents a potential therapeutical target in MM.
Assuntos
Mieloma Múltiplo/metabolismo , Proteínas de Neoplasias/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Idoso , Linhagem Celular Tumoral , Progressão da Doença , Intervalo Livre de Doença , Humanos , Mieloma Múltiplo/diagnóstico , Proteína de Sequência 1 de Leucemia de Células Mieloides , Valor Preditivo dos Testes , Recidiva , Estudos Retrospectivos , Resultado do TratamentoRESUMO
We have recently demonstrated that the CD40 molecule was expressed on both normal human plasma cells and most malignant plasma cells, i.e., myeloma cells. Thus, we have investigated its putative role in the proliferation of myeloma cells. We report that 7 of 15 myeloma cell lines were CD40+ but only one, XG2, presented a high level of CD40 expression. We show that the CD40 stimulation by anti-CD40 monoclonal antibodies (mAbs) of the interleukin 6-dependent myeloma cell line XG2 induced a total inhibition of its proliferation. This inhibition was also observed when cells were either cultured in the "CD40 system," where the anti-CD40 mAb has been immobilized on fibroblasts expressing Fc receptors or in the presence of a soluble chimeric CD40 ligand molecule. This inhibition of proliferation was neither accompanied by differentiation nor apoptosis. Triggering CD40 induced an homotypic aggregation of XG2 cells, and the inhibition of proliferation was totally prevented by a blocking anti-CD18 mAb. Although the CD11a-CD18 ligands, i.e., CD50, CD54, and CD102, were all expressed on XG2 cells, only a blocking anti-CD102 mAb inhibited the CD40-induced growth arrest. Our data demonstrate that CD40 triggering on XG2 cells induced a myeloma cell growth arrest mediated by lymphocyte function-associated antigen 1 and intercellular adhesion molecule 2 interactions.
Assuntos
Antígenos CD/fisiologia , Linfócitos B/imunologia , Antígenos CD11/fisiologia , Antígenos CD18/fisiologia , Antígenos CD40/fisiologia , Moléculas de Adesão Celular/fisiologia , Ciclo Celular , Divisão Celular/fisiologia , Anticorpos Monoclonais , Antígenos CD40/imunologia , Linhagem Celular , Citometria de Fluxo , Humanos , Imunoglobulina G , Interleucina-6/análise , Interleucina-6/biossíntese , Mieloma Múltiplo , Células Tumorais CultivadasRESUMO
In order to evaluate putative changes of major adhesion molecule expression on plasma cells (PCs) associated with malignant transformation, tumor spreading, and immortalization, we have quantified and compared the expression of CD56, CD44, CD11a, CD49e, and CD45 RO/RA on normal PCs, malignant PCs from multiple myeloma patients in chronic phase, in accelerated phase with or without extramedullary progression, and from human myeloma cell lines. Plasma cell phenotype was defined with the use of two-color immunofluorescence in combination with B-B4 or anti-CD38 antibodies. We found that all the adhesion antigens were expressed on normal PCs. Malignancy was characterized by an overexpression of CD56, whereas extramedullary spreading was associated with a dramatic down expression of CD56. Although CD44 remained unchanged, the subpopulation of PCs expressing CD11a, CD49e, and CD45RA/RO were significantly reduced during malignancy, and each of these negative subpopulations increased during disease acceleration. We demonstrated that CD11a and CD49e expression were correlated and defined the same subpopulation of PCs. The phenotype of HMCLs was similar to the expression profile of patients in accelerated phase with extramedullary spreading. In conclusion, we show that significant changes of PC phenotype were associated with malignancy, were correlated with the disease evolution, and could be of diagnostic and prognostic value in individuals with monoclonal gammopathy and patients with multiple myeloma.
Assuntos
Moléculas de Adesão Celular/metabolismo , Mieloma Múltiplo/patologia , Antígenos CD/metabolismo , Antígenos de Diferenciação de Linfócitos T/metabolismo , Medula Óssea/patologia , Antígeno CD56 , Proteínas de Transporte/metabolismo , Citometria de Fluxo , Humanos , Receptores de Hialuronatos , Técnicas In Vitro , Integrina alfa5 , Antígenos Comuns de Leucócito/metabolismo , Antígeno-1 Associado à Função Linfocitária/metabolismo , Plasmócitos/citologia , Plasmócitos/metabolismo , Receptores de Superfície Celular/metabolismo , Receptores de Retorno de Linfócitos/metabolismo , Células Tumorais CultivadasRESUMO
Dietary polyphenols constitute a large family of bioactive substances potential beneficial effect on metabolic syndrome (MetS). This review summarizes the results of clinical studies on patients with MetS involving the chronic supplementation of a polyphenol-rich diet, foods, extracts or with single phenolics on the features of MetS (obesity, dyslipidemia, blood pressure and glycaemia) and associated complications (oxidative stress and inflammation). Polyphenols were shown to be efficient, especially at higher doses, and there were no specific foods or extracts able to alleviate all the features of MetS. Green tea, however, significantly reduced body mass index and waist circumference and improved lipid metabolism. Cocoa supplementation reduced blood pressure and blood glucose. Soy isoflavones, citrus products, hesperidin and quercetin improved lipid metabolism, whereas cinnamon reduced blood glucose. In numerous clinical studies, antioxidative and anti-inflammatory effects were not significant after polyphenol supplementation in patients with MetS. However, some trials pointed towards an improvement of endothelial function in patients supplemented with cocoa, anthocyanin-rich berries, hesperidin or resveratrol. Therefore, diets rich in polyphenols, such as the Mediterranean diet, which promote the consumption of diverse polyphenol-rich products could be an effective nutritional strategy to improve the health of patients with MetS. © 2016 The Authors. Obesity Reviews published by John Wiley & Sons Ltd on behalf of World Obesity.
Assuntos
Antioxidantes/administração & dosagem , Dieta , Síndrome Metabólica/tratamento farmacológico , Polifenóis/administração & dosagem , Glicemia/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Suplementos Nutricionais , Humanos , Insulina/sangue , Resistência à Insulina , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
The present paper addresses the calibration of well-type ionization chambers (ICs) used at LNE-LNHB as standard transfer instruments to calibrate hospitals in the case of SIR-Spheres(®)(90)Y resin microspheres (Sirtex, Australia). Developed for interventional oncology, this radiopharmaceutical is directly injected in the liver for cancer treatment via a selective internal radiation therapy. The present work was carried out in the framework of the European project "Metrology for molecular radiotherapy" (MetroMRT). As commonly performed in radionuclide metrology for radiopharmaceuticals, the objective is to ensure the metrological traceability of SIR-Spheres(®) to hospitals. Preceding studies were focused on primary measurements of SIR-Spheres(®) based on the TDCR (Triple to Double Coincidence Ratio) method, applied after the dissolution of the (90)Y-labeled resin microspheres. As (90)Y is a high-energy ß(-)-emitter, the IC response strongly depends on the transport of electrons in the radioactive solution and surroundings (vial, chamber liners and materials). The variability of the IC-response due to the geometry dependence is investigated by means of measurements and Monte Carlo simulations in the case of a Vinten IC. The aim of the present study was also to propose a reliable uncertainty for ICs calibrations for the standard transfer of SIR-Spheres(®) to hospitals.
RESUMO
R115777, a nonpeptidomimetic farnesyl transferase inhibitor has recently demonstrated a significant antileukemic activity in vivo in acute myeloid leukemia. Multiple myeloma (MM) is a fatal hematological malignancy characterized by an accumulation of long-lived plasma cells within the bone marrow. In the present study, we have investigated the effect of the R115777 on growth and survival of myeloma cells. We have found that R115777 induced (1) a significant and dose-dependent growth inhibition of the three myeloma cell lines tested; and (2) a significant and time-dependent apoptosis. R115777 also induced apoptosis in the bone marrow mononuclear cell population of four MM patients, being almost restricted to the malignant plasma cells. Finally, we have investigated the effect of the R115777 in the Ras/MAPK and JAK/STAT pathways which are implicated in survival and/or proliferation in MM. The phosphorylation of both STAT3 and ERK1/2 induced by IL-6 was totally blocked at 15 microM of R115777 and partially blocked when R115777 was used at 10 and 5 microM. The induction of apoptosis by R115777 in myeloma cells and its implication in the regulation of JAK/STAT signalling suggest that R115777 might be an interesting therapeutical approach in MM.
Assuntos
Alquil e Aril Transferases/antagonistas & inibidores , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/patologia , Quinolonas/farmacologia , Idoso , Animais , Divisão Celular/efeitos dos fármacos , Proteínas de Ligação a DNA/antagonistas & inibidores , Farnesiltranstransferase , Feminino , Citometria de Fluxo , Humanos , Interleucina-6/metabolismo , Masculino , Pessoa de Meia-Idade , Proteína Quinase 1 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 3 Ativada por Mitógeno , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Prenilação de Proteína/efeitos dos fármacos , Fator de Transcrição STAT3 , Transativadores/antagonistas & inibidores , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
Multiple myeloma (MM) is a malignancy characterized by a very slow proliferation of malignant plasma cells leading to their accumulation within the bone marrow. This suggests that resistance to apoptosis may play a critical role both in the pathogenesis and resistance to treatment of MM. Bcl-2 is a key protein for the regulation of apoptosis. However, it has been shown that this protein also regulates the state of proliferation. In the current study, we show that malignant plasma cells from both the bone marrow and peripheral blood express high levels of Bcl-2 and are slowly proliferating cells. In contrast, myeloma cells from extramedullary sites (ie pleural effusion, ascitis, mammary and gastric plasmacytoma) express Bcl-2 weakly while being highly proliferative. Normal non-dividing bone marrow plasma cells express high levels of Bcl-2 protein. In contrast, four highly proliferative reactive plasmacytosis express weak levels of Bcl-2. We conclude that there is an inverse correlation between Bcl-2 expression and the proliferation rate of both normal and malignant plasma cells. These data may be explained by the double function of Bcl-2, ie its well known function as an anti-apoptotic molecule and its intriguing function as an inhibitory molecule of cell proliferation.
Assuntos
Leucemia Plasmocitária/metabolismo , Mieloma Múltiplo/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Adulto , Idoso , Células da Medula Óssea/patologia , Divisão Celular/fisiologia , Feminino , Humanos , Leucemia Plasmocitária/patologia , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/patologia , Plasmocitoma/metabolismo , Plasmocitoma/patologiaRESUMO
In this study, we show that malignant plasma cells from patients with either primary (n=12) or secondary (n=15) plasma cell leukemia (PCL) do not express CD56 at all, neither in the bone marrow nor the peripheral blood in 81% of cases. On the other hand, multiple myeloma (MM) at diagnosis overexpress it in 63 of 94 (67%) cases (P=0.0001). In three secondary PCL evaluated serially, CD56 was also lacking at diagnosis showing that CD56 is not downregulated at the end stage of the disease but rather not upregulated in this subset of patients. This last concept is strengthened by the observation that 29% of MM patients lacking CD56 or weakly expressing it at diagnosis present a detectable leukemic phase vs 11% only in CD561 MM (P=0.06). Forty percent of all the CD56(-/weak) malignant plasma cell disorders present or develop a leukemic phase vs only 15% of CD56+ cases (P < 0.008). CD56(-/weak) MM subset is also associated with a significantly less aggressive osteolytic potential (P=0.012). We conclude that the lack or weak expression of CD56 is a characteristic feature of PCL but also delineates a special subset of MM at diagnosis mainly characterized by a lower osteolytic potential and a trend for malignant plasma cells to circulate in the peripheral blood more overtly.
Assuntos
Antígeno CD56/metabolismo , Leucemia Plasmocitária/metabolismo , Mieloma Múltiplo/metabolismo , Proteínas de Neoplasias/metabolismo , Medula Óssea/metabolismo , Antígenos CD28/metabolismo , Diagnóstico Diferencial , Humanos , Leucemia Plasmocitária/diagnóstico , Mieloma Múltiplo/diagnóstico , Plasmócitos/metabolismoRESUMO
Multiple myeloma (MM) is a plasma-cell malignancy characterized by the accumulation of malignant plasma cells within the bone marrow. Interleukin (IL)-6 is an essential survival and growth factor for myeloma cells that exerts its activity through a cell surface receptor composed of an 80-kDa ligand binding molecule (IL-6Ralpha) and a 130-kDa signal-transducing molecule. Of major interest, the soluble form of the IL-6Ralpha (sIL-6Ralpha) is an agonistic molecule able to potentiate IL-6 activity and a strong prognostic factor in MM. In the present study, we demonstrate that purified myeloma cells from all of the patients with MM and human myeloma cell lines release sIL-6Ralpha. The level of sIL-6Ralpha release correlates with disease activity and is clearly up-regulated during tumoral expansion in vivo and immortalization in vitro. Of note, this sIL-6Ralpha release is strongly reduced (50%) by a hydroxamate-based metalloproteinase inhibitor underlying the importance of shedding in the production of sIL-6Ralpha by myeloma cells. Using specific IL-6Ralpha primers flanking the transmembrane domain, we demonstrate by PCR the presence of two IL-6R mRNAs corresponding to the membrane IL-6Ralpha and to the sIL-6Ralpha generated through alternative splicing in myeloma cells. In conclusion, we show that: (a) native myeloma cells and human myeloma cell lines release sIL-6Ralpha by two distinct mechanisms: alternative splicing and proteolytic cleavage of the membrane IL-6Ralpha; and (b) the release of the sIL-6Ralpha, which is an agonist of IL-6, correlates with disease progression, explaining in part its strong prognostic value in vivo.
Assuntos
Processamento Alternativo , Endopeptidases/fisiologia , Mieloma Múltiplo/metabolismo , Receptores de Interleucina-6/genética , Humanos , Mieloma Múltiplo/patologia , RNA Mensageiro/análise , Receptores de Interleucina-6/metabolismoRESUMO
CD28 expression was thoroughly investigated on plasma cells of monoclonal gammopathy of undetermined significance, multiple myeloma (MM), and human myeloma cell lines. CD28+ plasma cells were detected in 19% of 31 monoclonal gammopathy of undetermined significance, 41% of 116 MM, and 100% of 13 human myeloma cell lines. CD28+ myeloma cells were detected in 21 of 79 (26%) MM cases at diagnosis, 13 of 22 (59%) at medullary relapse (P < 0.009), and 14 of 15 (93%) at extramedullary relapse (P = 0.05), including 10 of 10 (100%) secondary plasma cell leukemias (P = 0.05). Serial studies in individual patients confirmed the emergence of CD28+ myeloma cells with tumoral expansion and treatment failure. This was significantly correlated with the expression of CD28 ligand, i.e., CD86 (but not CD80), and with an increase in the proliferative activity (labeling index) of myeloma cells in bone marrow. Whereas the expression of CD56 defines a particular subset of myeloma patients, CD28 is the only antigen for which expression correlates with tumor progression. Our data show that an aggressive compartment of CD28+ and CD86+ myeloma cells emerges during the course of MM in vivo, indicating that CD28 could be aberrantly expressed on highly malignant (possibly mutated) myeloma cells. Conversely, a subset of proliferative plasmablasts coexpressing CD28 and CD86 could be the normal counterpart of the clonogenic myeloma stem cell because a subset of CD28+ plasma cells was observed in 6 of 6 cases of reactive plasmocytosis.
Assuntos
Antígenos CD/análise , Biomarcadores Tumorais/análise , Medula Óssea/patologia , Antígenos CD28/análise , Mieloma Múltiplo/patologia , Antígeno B7-1/análise , Antígeno B7-2 , Antígeno CD56/análise , Linhagem Celular , Progressão da Doença , Humanos , Leucemia Plasmocitária/patologia , Glicoproteínas de Membrana/análise , Segunda Neoplasia Primária/patologia , Paraproteinemias/patologia , Valor Preditivo dos Testes , Recidiva , Falha de Tratamento , Células Tumorais CultivadasRESUMO
Half-life measurements have been carried out at LNHB for 88Y using a 4pi gamma-ionisation chamber and for 89Sr using a proportional counter. The determined half-life values and associated standard uncertainties are 106.63 +/- 0.05 d for 88Y and 50.65 +/- 0.05 d for 89Sr, being consistent with relevant values reported in literature. Based on the present results and relevant literature values revised recommended half-life values and associated standard uncertainties are proposed, viz. 106.626 +/- 0.021 d for 88Y and 50.57 +/- 0.03 d for 89Sr.