RESUMO
During tumor development, growth factors may act in autocrine manner stimulating cell proliferation, or in paracrine manner affecting the microenvironment of the tumor and modulating the immune system. Murine mammary adenocarcinoma M3 tumor bearers develop lung metastases and leukocytosis during its evolution. Previously we described that M3 conditioned media enhanced metastasis incidence, when it was inoculated in tumor-operated mice. In the present study we determine that spleen cells from M3 tumor operated mice treated with M3 conditioned media, were able to transfer the capacity to enhance metastasis to other tumor operated mice. Spleen cells have immune suppressor activity that could be reversed by cyclophosfamide treatment. M3 tumor cells secrete GM-CSF, which is able to promote in vitro proliferation of M3 cells as well as spleen cells. This proliferation could be abrogated by the addition of anti-GM-CSF. We report that the GM-CSF secreted by M3 tumor cells had stimulatory activity on M3 tumor cell and lymphocyte proliferation.
Assuntos
Adenocarcinoma/imunologia , Adenocarcinoma/metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Neoplasias Mamárias Animais/imunologia , Neoplasias Mamárias Animais/metabolismo , Adenocarcinoma/patologia , Animais , Antineoplásicos Alquilantes/farmacologia , Células da Medula Óssea/metabolismo , Divisão Celular , Células Cultivadas , Ensaio de Unidades Formadoras de Colônias , Meios de Cultivo Condicionados/farmacologia , Ciclofosfamida/farmacologia , Progressão da Doença , Feminino , Neoplasias Pulmonares/secundário , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Metástase Neoplásica , Baço/citologia , Baço/metabolismoRESUMO
The purpose of this study was to investigate if specific immune responses were present in mice bearing a lung adenocarcinoma that presents paraneoplastic syndromes during tumor evolution. Leukocytosis, mainly due to polymorphonuclear leukocytes, was found from day 15 of tumor growth. Delayed type hypersensitivity response and increased interleukin-6 (IL-6) serum levels were observed along tumor growth. Concomitant immunity, specific rejection of a second inoculum and in vitro specific cytotoxicity occurred at 20 days of implant. In advanced stages of tumor evolution impaired cytotoxicity, accompanied by a great increase of IL-6 in serum, were observed. Role of polymorphonuclear leukocytes and IL-6 overproduction as responsible for immune dysregulation and paraneoplastic syndromes are discussed.
Assuntos
Adenocarcinoma/imunologia , Neoplasias Pulmonares/imunologia , Síndromes Paraneoplásicas/imunologia , Adenocarcinoma/sangue , Animais , Divisão Celular , Hipersensibilidade Tardia/imunologia , Imunidade Celular , Imunização , Interleucina-6/sangue , Contagem de Leucócitos , Neoplasias Pulmonares/sangue , Linfócitos/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Síndromes Paraneoplásicas/sangue , Baço/patologia , Células Tumorais CultivadasRESUMO
CD44 is a widely distributed set of cell surface glycoproteins expressed in several types of cells and tissues, implicated in cell-cell and cell-substrate interactions. This molecule plays a major role in cell differentiation, development and activation and has also been described as a potential marker of malignancy and metastasis. In the present study we investigated by RT-PCR followed by exon specific amplification the expression of CD44 splice variants in four different murine tumors as well as in the invaded organs in order to correlate the expression of CD44 variants with potential tumor invasiveness and their implications for growth. Our data showed deregulation in the expression of CD44 isoforms but no discernible correlation in isoform expression pattern. However, in all tumors studied isoforms presented by the primary tumor were detected in the invaded organs before metastasis could be demonstrated by histopathological analysis.
Assuntos
Processamento Alternativo , Receptores de Hialuronatos/genética , Neoplasias/genética , Animais , Regulação Neoplásica da Expressão Gênica , Fígado/metabolismo , Pulmão/metabolismo , Linfonodos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias/patologia , Isoformas de Proteínas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Baço/metabolismo , Fatores de TempoRESUMO
LP07 is a new cell line derived from P07 lung tumor, spontaneously arisen in a BALB/c mouse. LP07 is composed of heterogeneous epithelioid polyhedric cells that proliferate at a slow rate, have low plating efficiency and are unable to grow in soft agar. Only some LP07 cells expressed cytokeratins while most of them were positive for vimentin. Ultrastructure studies showed that LP07 cells established rudimentary intercellular unions, formed glandular-like conducts and presented conspicuous secretory granules, suggesting an epithelial-glandular origin, with neuroendocrine components. Upon injection LP07 cells formed poorly differentiated non-invasive adenocarcinomas, and tumor bearing mice developed leukocytosis, hypercalcemia and cachexia. This tumor cell line constitutes a useful tool to study lung tumor biology and paraneoplastic syndromes.
Assuntos
Adenocarcinoma/patologia , Divisão Celular/fisiologia , Neoplasias Pulmonares/patologia , Síndromes Paraneoplásicas/patologia , Adenocarcinoma/sangue , Animais , Contagem de Células Sanguíneas , Peso Corporal , Cálcio/sangue , Testes de Carcinogenicidade , Adesão Celular , Linhagem da Célula , Movimento Celular , Cromossomos/genética , Análise Citogenética , Modelos Animais de Doenças , Feminino , Técnicas Imunoenzimáticas , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/secundário , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica , Transplante de Neoplasias , Síndromes Paraneoplásicas/sangue , Células Tumorais Cultivadas , Ativador de Plasminogênio Tipo Uroquinase/metabolismoRESUMO
Paraneoplastic syndromes are rarely described in animal models. It may be useful to have a suitable experimental model to study the mechanisms by which they are produced. In this study, we describe a murine lung adenocarcinoma, P07, which presents hypercalcemia, leukocytosis and cachexia. We determined the presence of PTHrP in plasma as well as GM-CSF produced by P07 cells. TNF-alpha, which is responsible for cachexia, could neither be detected in serum nor in P07 cell supernatants. We conclude that this model, which shows paraneoplastic syndromes similar to those of lung tumor patients, should be useful to study the pathways and significance of these signs.
Assuntos
Adenocarcinoma/fisiopatologia , Neoplasias Pulmonares/fisiopatologia , Síndromes Paraneoplásicas/fisiopatologia , Adenocarcinoma/sangue , Adenocarcinoma/patologia , Alanina Transaminase/sangue , Fosfatase Alcalina/sangue , Animais , Aspartato Aminotransferases/sangue , Proteínas Sanguíneas/análise , Caquexia/fisiopatologia , Cálcio/sangue , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/biossíntese , Hematócrito , Contagem de Leucócitos , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/patologia , Camundongos , Camundongos Endogâmicos BALB C , Síndromes Paraneoplásicas/sangue , Proteína Relacionada ao Hormônio Paratireóideo , Proteínas/análise , Células Tumorais CultivadasRESUMO
The design of more effective therapies for metastatic disease involves development of new compounds able to specifically block the malignant process. We demonstrated previously that a new synthetic nitrogenated compound 3'-1-chloroethyl-2,3-dihydro-1H-imidazo-(2, 1-i)-purine-4-ium-7-yl-3'-deoxy-1',5', 6'-tri-O-(methylsulfonyl)-muco-inositol chloride (DIC) had an anti-proliferative activity on tumor cells in vitro. In the present work we demonstrate that DIC induces apoptosis on the LM3 murine mammary adenocarcinoma cell line in vitro and has anti-angiogenic activity in vivo. We also evaluated toxicity, biodistribution and anti-neoplastic properties of DIC in vivo. Toxicity studies allowed us to establish the LD50 (750 mg/kg body weight). Administration of 250 mg/kg/day (LD10) for 6 days did not cause overt toxic effects. Biodistribution assays revealed that DIC was rapidly eliminated (60% at t=10 min), although it accumulated in tumor tissue at higher concentrations than in other tissues. Daily s.c. treatment with DIC (LD10) for 24 days significantly reduced the number of spontaneous lung metastases. These results suggest that DIC has the ability of impairing the metastatic development by inhibiting angiogenesis and inducing apoptosis on tumor cells.
Assuntos
Antineoplásicos/farmacologia , Inositol/análogos & derivados , Purinas/farmacologia , Adenocarcinoma/patologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/uso terapêutico , Antineoplásicos/toxicidade , Divisão Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Inositol/síntese química , Inositol/farmacologia , Inositol/toxicidade , Radioisótopos do Iodo , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Endogâmicos BALB C , Metástase Neoplásica/prevenção & controle , Neovascularização Patológica/prevenção & controle , Purinas/síntese química , Purinas/toxicidade , Distribuição Tecidual , Células Tumorais CultivadasRESUMO
We report the histological and biological behavior characteristics of a lung tumor (P07) that arose spontaneously in a Balb/c mouse. P07 is a moderately to poorly differentiated adenocarcinoma that secretes granulocyte-macrophage colony stimulating factor (GM-CSF) in culture supernatants. This tumor presents some paraneoplastic syndromes, such as leukocytosis, hypercalcemia and cachexia. taken together with the peripheral blood leukocyte (PBL) counts and serum calcium levels during s.c. tumor growth and after surgery, this study suggests that P07 may be a useful experimental model to study the biology of lung cancer and paraneoplastic syndromes.
Assuntos
Adenocarcinoma/fisiopatologia , Neoplasias Pulmonares/fisiopatologia , Síndromes Paraneoplásicas/fisiopatologia , Adenocarcinoma/patologia , Animais , Contagem de Células Sanguíneas , Peso Corporal , Medula Óssea/fisiologia , Cálcio/sangue , Modelos Animais de Doenças , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Hipercalcemia/etiologia , Leucocitose/etiologia , Neoplasias Pulmonares/patologia , Camundongos , Camundongos Endogâmicos BALB C , Síndromes Paraneoplásicas/patologiaRESUMO
We studied the effect of tumoral microenvironments on metastatic phenotypes. Therefore, murine mammary adenocarcinoma cells cultured in vivo in diffusion chambers (DC) were implanted intraperitoneally in BALB/c mice. The behavior of DC-cultured cells was compared with that of cells obtained from tumors growing subcutaneously or intraperitoneally and from primary cultures in vitro of the former. DC-cultured and control cells were inoculated into normal mice to evaluate their tumorigenicity and metastasizing ability. We found that DC-cultured cells were less tumorigenic and metastatic both in spontaneous and in experimental metastasis assays. The host response to tumor progression resulted in an early leukocytosis, probably due to the overproduction of a hematopoietic factor by the tumor cells. Finally, it was found that DC-cultured cells produced lower levels of urokinase-type plasminogen activator activity, while no differences were found in the metalloproteinase production compared to cells obtained from a tumor growing subcutaneously.
Assuntos
Adenocarcinoma/fisiopatologia , Neoplasias Mamárias Experimentais/fisiopatologia , Adenocarcinoma/patologia , Adenocarcinoma/secundário , Animais , Contagem de Células Sanguíneas , Cultura em Câmaras de Difusão/métodos , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/secundário , Neoplasias Mamárias Experimentais/patologia , Metaloendopeptidases/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Fenótipo , Células Tumorais Cultivadas , Ativador de Plasminogênio Tipo Uroquinase/biossínteseRESUMO
A role for nitric oxide (NO) in the regulation of blood leukocyte numbers was examined in BALB/c mice by employing the NO synthase inhibitor NG-nitro L-arginine methyl ester (L-NAME). Treatment of animals with a single dose of 50 mg/kg body wt caused a dramatic increase in the number of circulating neutrophils and a moderate decrease in the number of circulating lymphocytes. These effects were partially reversed by the simultaneous inoculation of L-arginine (250 mg/kg body wt.) but not by D-arginine. A second NO synthase inhibitor, NG-nitro L-arginine, induced changes comparable to those elicited by L-NAME. Because catecholamines and glucocorticoids are well-known modulators of blood leukocyte counts, experiments were carried out in adrenalectomized mice. It was found that adrenalectomy did not modify the increase in the number of circulating neutrophils induced by L-NAME but completely prevented the decrease of circulating lymphocytes. Taken together, these findings support the hypothesis that NO plays an important role in the regulation of the peripheral blood number of neutrophils and lymphocytes, and that this function involves, in each case, the participation of different mechanisms.
Assuntos
Linfócitos/citologia , Óxido Nítrico/fisiologia , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Inibidores Enzimáticos/farmacologia , Contagem de Leucócitos/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Linfopenia/sangue , Linfopenia/induzido quimicamente , Masculino , Camundongos , Camundongos Endogâmicos BALB C , NG-Nitroarginina Metil Éster , Neutropenia/sangue , Neutropenia/induzido quimicamente , Neutrófilos/citologia , Neutrófilos/efeitos dos fármacos , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico Sintase/antagonistas & inibidores , Nitroarginina , EstereoisomerismoRESUMO
We have studied in BALB/c mice the hematological alterations of the host induced by the growth of tumor cells in diffusion chambers (DC) In this model, host-tumor interactions are only mediated by soluble factors. Tumor cells proliferate and grow in DC up to 15 days after implant. Our results show a reversal of the granulocyte-lymphocyte ratio in peripheral blood, with lymphopenia and a relative increase of myeloid progenitors in the bone marrow of mice bearing DC with M3 tumor cells (M3TC).