RESUMO
The multidrug resistance (MDR) transporter-proteins P-glycoprotein (Pgp), multidrug resistance protein (MRP) and lung resistance protein (LRP) have been associated with treatment failure. The aim of this study was to investigate prospectively the clinical significance of expression and function of the MDR proteins, considering other prognostic factors, such as age, immunophenotype, and cytogenetics. Mononuclear cells of peripheral blood or bone marrow from 61 patients with de novo acute myelogenous leukemia (AML) were analyzed. The monoclonal antibodies JSB1, MRPm6 and LRP56 were used for expression studies. Accumulation and retention studies were performed using the substrates Daunorubicin, Calcein-AM, Rhodamine-123 and DiOC(2) in the presence or absence of the modifiers Verapamil, Genistein, Probenecid, BIBW22S and PSC833. Induction treatment consisted of a 3+7 combination of Ida/Ara-C for patients < or = 60 years of age and a 3+5 Ida/VP-16 combination per OS for patients >60. MDR function was expressed as the ratio of mean fluorescence intensity substrate in the presence of modifier over the substrate alone (resistance index, RI). Patients with advanced age, low CD15 expression and high RI for accumulation of DiOC(2) in the presence of BIBW22S had significantly lower complete remission (CR) rates. No factor was prognostic for event-free survival analysis, which was limited to remitters only. Overall survival was shorter in patients with advanced age, poor prognosis cytogenetics, high CD7 expression, and high RI for Daunorubicin efflux modulated by Verapamil. These results suggest that MDR transporter-proteins have a limited role in the treatment failure of patients treated with Idarubicin-based regimens.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/fisiologia , Resistência a Múltiplos Medicamentos/genética , Resistencia a Medicamentos Antineoplásicos/genética , Leucemia Mieloide/metabolismo , Proteínas Associadas à Resistência a Múltiplos Medicamentos/fisiologia , Proteínas de Neoplasias/fisiologia , Partículas de Ribonucleoproteínas em Forma de Abóbada/fisiologia , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/biossíntese , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Doença Aguda , Adolescente , Adulto , Fatores Etários , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Transplante de Medula Óssea , Bloqueadores dos Canais de Cálcio/farmacologia , Carbocianinas/metabolismo , Terapia Combinada , Citarabina/administração & dosagem , Daunorrubicina/metabolismo , Intervalo Livre de Doença , Inibidores Enzimáticos/farmacologia , Feminino , Fluoresceínas/metabolismo , Corantes Fluorescentes/metabolismo , Genisteína/farmacologia , Humanos , Idarubicina/administração & dosagem , Imunofenotipagem , Leucemia Mieloide/tratamento farmacológico , Leucemia Mieloide/genética , Leucemia Mieloide/mortalidade , Leucemia Mieloide/terapia , Masculino , Pessoa de Meia-Idade , Proteínas Associadas à Resistência a Múltiplos Medicamentos/biossíntese , Proteínas Associadas à Resistência a Múltiplos Medicamentos/genética , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética , Probenecid/farmacologia , Prognóstico , Estudos Prospectivos , Rodamina 123/metabolismo , Análise de Sobrevida , Células Tumorais Cultivadas/metabolismo , Partículas de Ribonucleoproteínas em Forma de Abóbada/biossíntese , Partículas de Ribonucleoproteínas em Forma de Abóbada/genética , Verapamil/farmacologiaRESUMO
Coexistence of Philadelphia chromosome-negative (Ph-) progenitors with the Ph+ clone in the early chronic phase of chronic myeloid leukemia (CML) has been documented in previous reports. A different evaluation of methods is needed to justify the clonality of the residual Ph- progenitors. Therefore, the X chromosome inactivation patterns in individual granulocyte-monocyte colony-forming unit (CFU-GM) colonies were studied with the clonality assay for the human androgen receptor gene. A prerequisite for this evaluation was the validation of T-lymphocytes and buccal cells as control cells representing the constitutional lyonization. The percentages of polyclonal CFU-GM cells were determined in 9 Ph+ women with CML and in 5 healthy women. Results of the clonal analysis of CFU-GM colonies were compared with those from reverse transcriptase-polymerase chain reaction analysis of single colonies for BCR/ABL transcripts. Both methods of CFU-GM cell analysis were in agreement regarding the presence of variable proportions (0%-94%) of normal cells in CML. Our results suggest that (a) T-cells and buccal cells have potential for use as controls for the clonal analysis of CML cases and (b) this method can evaluate the frequency of polyclonal/clonal CFU-GM cells in CML cases and is applicable to the analysis of myeloid clonal disorders that lack specific molecular markers.
Assuntos
Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Células Progenitoras Mieloides/patologia , Receptores Androgênicos/genética , Adulto , Estudos de Casos e Controles , Células Clonais/patologia , Mecanismo Genético de Compensação de Dose , Feminino , Proteínas de Fusão bcr-abl/genética , Humanos , Leucemia Mieloide Crônica Atípica BCR-ABL Negativa/patologia , Métodos , Pessoa de Meia-Idade , RNA Mensageiro/análiseRESUMO
Polycythemia vera (PV) and essential thrombocythemia (ET) are chronic myeloproliferative diseases that carry intrinsically the potential for leukemic transformation. The aims of this study were (1) to detect involvement of N- and K-ras mutations in codons 12 and 13 in the pathogenesis of the chronic and blastic phases of PV and ET, (2) to study the occurrence of microsatellite instability (MSI) in chromosomes 5 and 7 during the chronic phase and blastic transformation of the disease, and (3) to examine the incidence of leukemia in patients treated with hydroxyurea (HU). Samples of PV and ET patients were analyzed with a polymerase chain reaction. No N- or K-ras mutations were detected. A positive score for MSI in chromosome 7 was found in 1 patient with PV during leukemic transformation. Three of 69 patients developed acute myelogenous leukemia, 2 with PV and 1 with ET. As of this report, the overall incidence of leukemic transformation is 5.7% (2/35 patients) in PV and 3.3% (1/30 patients) in ET patients treated with HU. These results indicate that (1) MSI is a genetic marker that can be detected, even in a small group of patients, at the blastic phase of the disease and (2) no increased leukemogenicity was noted in this group of patients treated with HU.
Assuntos
Cromossomos Humanos Par 5/genética , Cromossomos Humanos Par 7/genética , Hidroxiureia/efeitos adversos , Leucemia/induzido quimicamente , Policitemia Vera/complicações , Trombocitose/complicações , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Transformação Celular Neoplásica/induzido quimicamente , Transformação Celular Neoplásica/genética , Cromossomos Humanos Par 5/fisiologia , Cromossomos Humanos Par 7/fisiologia , Feminino , Genes ras/genética , Genes ras/fisiologia , Humanos , Hidroxiureia/uso terapêutico , Incidência , Leucemia/etiologia , Leucemia/genética , Masculino , Repetições de Microssatélites , Pessoa de Meia-Idade , Mutação , Policitemia Vera/tratamento farmacológico , Policitemia Vera/genética , Trombocitose/tratamento farmacológico , Trombocitose/genéticaRESUMO
The information concerning potential effects of somatic hypermutation on bcl-2 sequences translocated to the immunoglobulin heavy chain gene (IgH) locus in follicular lymphoma (FL) is rather limited. We analysed the complete open reading frame (ORF) of the bcl-2 gene for the presence of mutations in 24 bcl-2/IgH-positive diagnostic FL samples by the single strand conformation polymorphism (SSCP) technique. A prior analysis on many of these FL samples had revealed a consistent pattern of somatic hypermutation in IgH genes. Abnormally migrating bands on SSCP gels were identified only in 4/24 samples. This result provides strong support for the notion that in FL the translocated bcl-2 coding region is not targeted by somatic hypermutation.
Assuntos
Cromossomos Humanos Par 14 , Cromossomos Humanos Par 18 , Genes bcl-2/genética , Linfoma Folicular/genética , Hipermutação Somática de Imunoglobulina , Translocação Genética , Análise Mutacional de DNA , Rearranjo Gênico de Cadeia Pesada de Linfócito B/genética , Humanos , Linfoma Folicular/patologia , Fases de Leitura Aberta/genéticaAssuntos
Leucemia Megacarioblástica Aguda , Policitemia Vera/patologia , Sobreviventes , Medula Óssea/patologia , Terapia Combinada , Progressão da Doença , Transfusão de Eritrócitos , Feminino , Humanos , Hidroxiureia/uso terapêutico , Interferon-alfa/uso terapêutico , Leucemia Megacarioblástica Aguda/patologia , Policitemia Vera/tratamento farmacológico , Mielofibrose Primária/patologiaRESUMO
We report a patient with chronic myelogenous leukemia in chronic phase and basophilia which was found to carry a simple variant t(16;22) (q24;q11) Philadelphia (Ph) chromosome in unstimulated bone marrow mononuclear cells. Molecular analysis of peripheral blood and bone marrow mononuclear cells demonstrated the presence of a bcr-abl chimeric mRNA transcript of the b(3) -a(2) type. These findings confirm that band 9q34 participates in the formation of all Ph chromosomes, either standard or variant, even when this is not detectable by conventional cytogenetics. The available literature concerning variant Philadelphia translocations is also reviewed.
RESUMO
BACKGROUND AND OBJECTIVES: Early lymphoid differentiation is characterized by antigen receptor gene rearrangements; the rearrangement process is governed by two lymphoid-specific genes, RAG (recombinase activating gene)-1 and -2. The available data on the incidence and prognostic significance of clonal immunoglobulin heavy chain (IgH) gene rearrangements in acute myeloid leukemia (AML) are rather contradictory. The aim of this study was to evaluate the incidence and prognostic significance of RAG-1 and -2 mRNA transcripts and clonal IgH gene rearrangements in a cohort of uniformly treated AML patients; the available literature is also reviewed. DESIGN AND METHODS: The study was performed on 76 AML patients, newly diagnosed between August 1996 and November 1999. RAG-1/-2 gene expression was analyzed by a reverse transcriptase polymerase chain reaction technique and IgH gene rearrangements were detected with variable region (VH) family-specific and consensus framework region (FWR)-2 and/or-3 primers. Statistical associations were explored between IgH monoclonality/ RAG mRNA expression and: (i) age, gender, FAB subtype, immunophenotype, cytogenetic risk groups; (ii) response variables (response/relapse incidence, survival). RESULTS: In total, 38/75 samples (50.6%) were RAG-1 and/or -2 positive; 30/76 samples (39.5%) carried clonal IgH genes, whereas 13/30 IgH-positive samples (43.3%) were RAG-1/2-negative. Significant associations were detected only for RAG-2 positivity and unfavorable karyotype and IgH monoclonality and FAB subtypes M4/ M5; no association was identified with response outcome and survival. INTERPRETATION AND CONCLUSIONS: Lymphoid-specific molecular markers are detected in a significant proportion of AML patients, regardless of differentiation status (assessed morphologically/ immunophenotypically); however, in our experience, they do not seem to constitute an adverse prognostic factor.
Assuntos
Proteínas de Ligação a DNA/genética , Regulação Neoplásica da Expressão Gênica , Genes de Imunoglobulinas/genética , Genes RAG-1/genética , Leucemia Mieloide/genética , Doença Aguda , Biomarcadores , Medula Óssea , Feminino , Rearranjo Gênico , Humanos , Leucemia Mieloide/diagnóstico , Leucemia Mieloide/mortalidade , Masculino , Pessoa de Meia-Idade , Proteínas Nucleares , Prognóstico , RNA Mensageiro/análise , Taxa de SobrevidaRESUMO
Interaction between receptor activator of nuclear factor kappaB ligand (RANKL) and RANK/osteoprotegerin (OPG) plays a dominant role in osteoclast activation and possibly in plasma cell survival in multiple myeloma (MM). We measured soluble RANKL (sRANKL), OPG, and bone remodeling markers in 121 patients with newly diagnosed MM to evaluate their role in bone disease and survival. Serum levels of sRANKL were elevated in patients with MM and correlated with bone disease. The sRANKL/OPG ratio was also increased and correlated with markers of bone resorption, osteolytic lesions, and markers of disease activity. The sRANKL/OPG ratio, C-reactive protein (CRP), and beta2-microglobulin were the only independent prognostic factors predicting survival in multivariate analysis. We generated a prognostic index based on these factors that divided our patients into 3 risk groups. The low-risk group had a 96% probability of survival at 5 years, whereas the intermediate-risk and the high-risk groups had probabilities of survival of 52% and 0%, respectively. Not only do these results confirm for the first time in humans the importance of sRANKL/OPG in the development of bone disease, they also highlight the role of this pathway in the biology of plasma cell growth as reflected by its influence on survival.
Assuntos
Proteínas de Transporte/sangue , Glicoproteínas/sangue , Glicoproteínas de Membrana/sangue , Mieloma Múltiplo/patologia , Receptores Citoplasmáticos e Nucleares/sangue , Índice de Gravidade de Doença , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Doenças Ósseas/diagnóstico , Doenças Ósseas/etiologia , Remodelação Óssea , Proteína C-Reativa/análise , Sobrevivência Celular , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/diagnóstico , Mieloma Múltiplo/mortalidade , Análise Multivariada , Osteoprotegerina , Prognóstico , Ligante RANK , Receptor Ativador de Fator Nuclear kappa-B , Receptores do Fator de Necrose Tumoral , Solubilidade , Análise de Sobrevida , Microglobulina beta-2/sangueRESUMO
Tartrate-resistant acid phosphatase isoform-5b (TRACP-5b), a new marker reflecting osteoclast activity, and osteoprotegerin (OPG) were measured in 121 patients with multiple myeloma (MM) at diagnosis, and in 63 of them during pamidronate administration, to define their correlation with the extent of bone disease and disease activity in MM. Radiographic evaluation of the skeleton, measurement of other markers of bone remodelling, including N-terminal cross-linking telopeptide of type-I collagen (NTX), bone alkaline phosphatase and osteocalcin and of markers of disease activity (beta2-microglobulin, paraprotein, interleukin-6 (IL-6), were also performed. Levels of TRACP-5b were increased (p <.0001), while OPG was decreased in MM patients compared to controls (p <.01). TRACP-5b levels were associated with the radiographically assessed severity of bone disease (p <.0001) as well as with levels of NTX, IL-6 and beta2-microglobulin (p <.001, for each biochemical parameter, respectively). The combination of pamidronate with VAD-chemotherapy produced a reduction in TRACP-5b, NTX, IL-6, paraprotein and beta2-microglobulin levels from the 2nd month of treatment, with no effect on bone formation and OPG. A strong correlation was observed between changes in TRACP-5b and changes in NTX, IL-6 and beta2-microglobulin, while TRACP-5b predicted the disease progression in 5 patients. These findings suggest that TRACP-5b is increased in MM, reflects the extent of myeloma bone disease and may have a predictive value. TRACP-5b has also proved to be very useful for monitoring antimyeloma treatment, which had no effect on OPG levels.
Assuntos
Fosfatase Ácida/sangue , Biomarcadores Tumorais/sangue , Biomarcadores/análise , Neoplasias Ósseas/enzimologia , Isoenzimas/sangue , Mieloma Múltiplo/enzimologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Fosfatase Alcalina/sangue , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias Ósseas/tratamento farmacológico , Proteína C-Reativa/metabolismo , Citarabina/uso terapêutico , Dexametasona/uso terapêutico , Difosfonatos/uso terapêutico , Feminino , Glicoproteínas/metabolismo , Humanos , Interleucina-6/sangue , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/tratamento farmacológico , Osteocalcina/sangue , Osteoclastos/enzimologia , Osteoclastos/metabolismo , Osteogênese/efeitos dos fármacos , Osteoprotegerina , Pamidronato , Paraproteínas/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Receptores do Fator de Necrose Tumoral , Valores de Referência , Sensibilidade e Especificidade , Fosfatase Ácida Resistente a Tartarato , Vincristina/uso terapêutico , Microglobulina beta-2/sangueRESUMO
BACKGROUND: Postremission therapy is critical in maintaining complete remission (CR) in patients with de novo acute myelogenous leukemia (AML). The aim of this trial was to compare allogeneic stem cell transplantation (SCT), high-dose cytarabine (ara-C; HiDAC), and autologous SCT as postremission therapy in patients with de novo AML. METHODS: One hundred twenty patients age = 60 years with previously untreated AML (non-M3) and a performance status score of = 2 received induction therapy with 3 days of idarubicin and 7 days of ara-C (IA). Patients in CR received one course of HiDAC. Subsequently, patients age = 50 years with available HLA-compatible donors were assigned to receive allogeneic SCT; patients with "favorable" cytogenetics received a second course of HiDAC; and all others were randomized to a second course of HiDAC or autologous SCT. RESULTS: The IA combination induced CR in 99 patients (82.5%). With a median follow-up of 43 months (range, 18-64 years), the 3-year survival and failure-free survival (FFS) rates were 47% and 45%, respectively. The factors associated with longer survival were those identified for CR (i.e., age and cytogenetics). Forty-nine patients (49%) received the assigned postremission therapy. Fifteen patients underwent allogeneic SCT. Nineteen patients underwent autologous SCT and 15 patients received a second course of HiDAC, after randomization. In the allogeneic SCT group, both the 3-year survival and the FFS rates were 73%. In the autologous SCT and HiDAC groups, the 3-year survival rates were 58% and 46%, respectively (P = 0.80), and the 3-year FFS rates were 42% and 33%, respectively (P = 0.83). CONCLUSIONS: The three postremission treatment groups had comparable survival. Allogeneic SCT is associated with a prolonged FFS.
Assuntos
Citarabina/administração & dosagem , Leucemia Mieloide Aguda/terapia , Transplante de Células-Tronco , Adolescente , Adulto , Esquema de Medicação , Feminino , Humanos , Leucemia Mieloide Aguda/mortalidade , Masculino , Pessoa de Meia-Idade , Indução de Remissão , Análise de Sobrevida , Condicionamento Pré-Transplante , Transplante Autólogo , Transplante Homólogo , Resultado do TratamentoRESUMO
OBJECTIVES: Bisphosphonates have been found to reduce skeletal events in patients with multiple myeloma (MM). This is the first randomised trial to compare the efficacy of pamidronate and ibandronate, a third-generation aminobisphosphonate, in bone turnover and disease activity in MM patients. METHODS: Patients with MM, stage II or III, were randomly assigned to receive either pamidronate 90 mg (group I: 23 patients) or ibandronate 4 mg (group II: 21 patients) as a monthly intravenous infusion in addition to conventional chemotherapy. Skeletal events, such as pathologic fractures, hypercalcaemia, and bone radiotherapy were analysed. Bone resorption markers [N-terminal cross-linking telopeptide of type-I collagen (NTX) and tartrate-resistant acid phosphatase type 5b (TRACP-5b)], bone formation markers (bone alkaline phosphatase and osteocalcin), markers of disease activity (paraprotein, CRP, beta 2-microglobulin), and interleukin-6 (IL-6) were also studied. RESULTS: In both groups, the combination of chemotherapy with either pamidronate or ibandronate produced a reduction in bone resorption and tumour burden as measured by NTX, IL-6, paraprotein, CRP, and beta 2-microglobulin from the second month of treatment, having no effect on bone formation. TRACP-5b also had a significant reduction in the pamidronate group from the second month of treatment and in the ibandronate group from the sixth month. However, there was a greater reduction of NTX, IL-6, and beta 2-microglobulin in group I than in group II, starting at the second month of treatment (P = 0.002, 0.001, and 0.004, respectively) and of TRACP-5b, starting at the fourth month (P = 0.014), that being continued throughout the 10-month follow-up of this study. There was no difference in skeletal events during this period. A significant correlation was observed between changes of NTX and changes of TRACP-5b, IL-6, and beta 2-microglobulin from the second month for patients of both groups. CONCLUSIONS: These results suggest that a monthly dose of 90 mg of pamidronate is more effective than 4 mg of ibandronate in reducing osteoclast activity, bone resorption, IL-6, and possibly tumour burden in MM. TRACP-5b has also proved to be a useful new marker for monitoring bisphosphonates treatment in MM.
Assuntos
Reabsorção Óssea/tratamento farmacológico , Difosfonatos/administração & dosagem , Mieloma Múltiplo/tratamento farmacológico , Fosfatase Ácida/sangue , Idoso , Antineoplásicos/administração & dosagem , Biomarcadores/sangue , Biomarcadores/urina , Reabsorção Óssea/etiologia , Reabsorção Óssea/prevenção & controle , Colágeno/urina , Colágeno Tipo I , Feminino , Humanos , Ácido Ibandrônico , Interleucina-6/sangue , Isoenzimas/sangue , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/complicações , Osteogênese/efeitos dos fármacos , Pamidronato , Peptídeos/urina , Fosfatase Ácida Resistente a Tartarato , Microglobulina beta-2/sangueRESUMO
Treatment with recombinant human erythropoietin (rHuEpo) improves anaemia in approximately 20% of patients with myelodysplastic syndromes (MDS). We investigated the potential advantage of a prolonged administration of rHuEpo to achieve higher erythroid response rates (RR) in 281 MDS patients: 118 with refractory anaemia (RA), 77 with refractory anaemia and ringed sideroblasts (RARS), 59 with refractory anaemia with excess of blasts and blast count < 10% (RAEB-I), and 27 with RAEB and blast count between 11-20% (RAEB-II). rHuEpo was given subcutaneously at a dose of 150 U/kg thrice weekly, for a minimum of 26 weeks. Response to treatment was evaluated after 12 and 26 weeks of therapy. The overall RR was 45.1%; the RR for RA, RARS, RAEB-I and RAEB-II were 48.3%, 58.4%, 33.8% and 13% respectively. A significant increase in RR was observed at week 26 in RA, RARS and RAEB-I patients, as the response probability increased with treatment duration. The RR was higher in the good cytogenetic prognostic group and serum Epo level of > 150 U/l at baseline predicted for non-response. The median duration of response was 68 weeks and the overall risk of leukaemic transformation was 21.7%. These results suggest that prolonged administration of rHuEpo produces high and long-lasting erythroid RR in MDS patients with low blast counts, particularly in those with pretreatment serum Epo levels of < 150 U/l and good cytogenetic prognosis.