RESUMO
SPARC (secreted protein acidic and rich in cysteine) is a matricellular protein highly expressed during development, reorganization and tissue repair. In the central nervous system, glial cells express SPARC during development and in neurogenic regions of the adult brain. Astrocytes control the glutamate receptor levels in the developing hippocampus through SPARC secretion. To further characterize the role of SPARC in the brain, we analyzed the hippocampal-dependent adult behavior of SPARC KO mice. We found that SPARC KO mice show increased levels of anxiety-related behaviors and reduced levels of depression-related behaviors. The antidepressant-like phenotype could be rescued by adenoviral vector-mediated expression of SPARC in the adult hippocampus, but anxiety-related behavior persisted in these mice. To identify the cellular mechanisms underlying these behavioral alterations, we analyzed neuronal activity and neurogenesis in the dentate gyrus (DG). SPARC KO mice have increased levels of neuronal activity, evidenced as more neurons that express c-Fos after a footshock. SPARC also affects cell proliferation in the subgranular zone of the DG, although it does not affect maturation and survival of new neurons. SPARC expression in the adult DG does not revert the proliferation phenotype in KO mice, but our results suggest a role of SPARC in limiting the survival of new neurons in the DG. This work suggests that SPARC could affect anxiety-related behavior by modulating neuronal activity, and that depression-related behavior is dependent upon the adult expression of SPARC, which affects adult brain function by mechanisms that need to be elucidated.
Assuntos
Depressão/genética , Hipocampo/fisiopatologia , Osteonectina/genética , Fatores Etários , Animais , Ansiedade/genética , Ansiedade/fisiopatologia , Proliferação de Células , Giro Denteado/fisiopatologia , Depressão/fisiopatologia , Feminino , Masculino , Camundongos , Camundongos Knockout , Neurogênese/genética , FenótipoRESUMO
Mesenchymal stem (stromal) cells (MSCs) are a source of circulating progenitors that are able to generate cells of all mesenchymal lineages and to cover cellular demands of injured tissues. The extent of their transdifferentiation plasticity remains controversial. Cells with MSC properties have been obtained from diverse tissues after purification and expansion in vitro. These cellular populations are heterogeneous and under certain conditions show pluripotent-like properties. MSCs present immunosuppressive and anti-inflammatory features and high migratory capacity toward inflamed or remodeling tissues. In this study we review available data regarding factors and signaling axes involved in the chemoattraction and engraftment of MSCs to an injured tissue or to a tissue undergoing active remodeling. Moreover, experimental evidence in support of uses of MSCs as vehicles of therapeutic genes is discussed. Because of its regenerative capacity and its particular immune properties, the liver is a good model to analyze the potential of MSC-based therapies. Finally, the potential application of MSCs and genetically modified MSCs in liver fibrosis and hepatocellular carcinoma (HCC) is proposed in view of available evidence.
Assuntos
Carcinoma Hepatocelular/terapia , Terapia Genética , Cirrose Hepática/terapia , Neoplasias Hepáticas/terapia , Células-Tronco Mesenquimais , Animais , Quimiotaxia , Técnicas de Transferência de Genes , Engenharia Genética , Células-Tronco Mesenquimais/imunologia , Células-Tronco Mesenquimais/fisiologia , CamundongosRESUMO
Cell-based gene therapy after cytokine gene transfer is being investigated for autologous and allogeneic vaccination in cancer therapy. Here we show that mice vaccinated with 3-5 x 10(6) interleukin 12 (IL-12) gene-transduced CT26 colon cancer cells developed a long-lasting antitumor immune memory able to reject not only parental cells but also syngeneic, LM3 mammary, and MCE fibrosarcoma tumorigenic cells. In contrast, mice vaccinated with 0.5-1 x 10(6) CT26 cells transduced with pBabe neo IL-12 retrovirus cells (CT26-IL12) were only able to reject parental cells. An increase in the total circulating levels of IgG2a and a clear shift toward a systemic Th1 response developed, regardless of the amount of injected CT26-IL12 cells. On the contrary, a strong increase in anti-CT26-specific IgG2a levels was observed only when 3-5 x 10(6) CT26-IL12 cells were injected. Immunocompetent mice vaccinated with 3-5 x 10(6) CT26-IL12 cells developed local nodules for a few days, which then ceased growing. These nodules comprised mainly blood vessels, suggesting that an angiogenic process was taking place. CD8+ T cells were responsible for the anti-LM3 tumor cell memory, whereas CD4+ T cells were not involved. Splenocytes and lymphocytes obtained from mice immunized against CT26 cells were able to kill LM3 cells in vitro. Adoptive transfer of lymphocytes obtained from animals immunized against CT26 colon cancer cells suppressed LM3 mammary tumor growth in tumor-bearing mice. The present studies raised the possibility of isolating CTL clones and identifying CTL epitopes shared by different tumor cell types, which can be a target for cancer therapy.
Assuntos
Vacinas Anticâncer/imunologia , Neoplasias do Colo/imunologia , Neoplasias do Colo/terapia , Fibrossarcoma/terapia , Interleucina-12/imunologia , Neoplasias Mamárias Experimentais/terapia , Animais , Especificidade de Anticorpos , Linfócitos T CD8-Positivos/imunologia , Vacinas Anticâncer/genética , Vacinas Anticâncer/uso terapêutico , Neoplasias do Colo/genética , Neoplasias do Colo/prevenção & controle , Fibrossarcoma/imunologia , Fibrossarcoma/prevenção & controle , Técnicas de Transferência de Genes , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Memória Imunológica/imunologia , Imunoterapia Adotiva/métodos , Interleucina-12/genética , Linfócitos/imunologia , Neoplasias Mamárias Experimentais/imunologia , Neoplasias Mamárias Experimentais/prevenção & controle , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neovascularização Patológica/imunologia , Células Th1/imunologia , Transdução GenéticaRESUMO
Competitive PCR is a highly sensitive method for specific DNA quantification. Despite the lack of studies related to the accuracy of the method it has been widely used. Here we present a simulation model for competitive PCR, which takes into account the efficiency decay as a linear relationship of the total product yield. The model helped us to study the kind and magnitude of errors that arise from quantitative and semiquantitative competitive PCR protocols and to find ways to minimize them. The simulation data suggest that differences in amplification efficiency between target and standard templates induce stronger biases in quantitative than in semiquantitative competitive PCR. Quantitative competitive PCR can only be used when both efficiencies are equal. In contrast, semiquantitative competitive PCR can be used even when the target is amplified with a higher efficiency than the standard, since under such conditions the method tends to underestimate the differences in initial DNA content. These predictions have been confirmed with experimental data and show that the estimation of the amplification efficiencies is a prerequisite for the use of quantitative and semiquantitative competitive PCR. A simple method for this estimation is also presented.
Assuntos
DNA/análise , Reação em Cadeia da Polimerase/métodos , Microglobulina beta-2/genética , Animais , Primers do DNA/química , Eletroforese em Gel de Poliacrilamida , Processamento de Imagem Assistida por Computador , Ratos , Padrões de Referência , Reprodutibilidade dos Testes , Microglobulina beta-2/metabolismoRESUMO
Tumor cells transduced with retrovirus carrying the herpes simplex-1 virus thymidine kinase (HSV-tk) are capable of transforming the antiviral drug ganciclovir (GVC) into a metabolic form only toxic to dividing cells. The efficiency of this suicide gene therapy is increased by a "bystander" effect resulting not only in the death of the recipient cell, but also in the death of non modified surrounding cells. Even though the mechanism of this "bystander" effect remains to be elucidated, strong evidence suggest that the immune system plays a main role to achieve complete tumor eradication. In the present study we evaluate the efficiency of this suicide system on three different tumor models: one human melanoma, one murine melanoma, and a rat glioblastoma. Tumors were established by injection of tumor cells s.c. in nude and C57Bl/6 mice, respectively, and stereotactically into the brain of Sprague Dawley rats. Animals in the treated group were co-injected with packaging cells producing recombinant retrovirus carrying the HSV-tk gene, and followed by i.p. administration of GVC. In short term studies, we observed inhibition of tumor growth for all the tumor models evaluated (p < 0.01). In long term studies, using the C6 rat glioma line, 50% of the animals survived longer than 75 days (p < 0.0001), and were able to reject a contralateral challenges with C6 parental cells. Histological and immunohistochemical analysis showed the presence at an inflammatory infiltrate composed by T lymphocytes, macrophages and polymorphonuclear cells. These data demonstrate that suicide genes might represent an attractive form of cancer gene therapy in the treatment of brain tumors and their intracerebral dissemination.
Assuntos
Antimetabólitos/farmacologia , Neoplasias Encefálicas/terapia , Ganciclovir/farmacologia , Técnicas de Transferência de Genes , Terapia Genética/métodos , Glioma/terapia , Melanoma Experimental/terapia , Timidina Quinase/genética , Animais , Encéfalo/patologia , Morte Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Vetores Genéticos , Herpesvirus Humano 1/genética , Camundongos , RatosRESUMO
SPARC (secreted protein acidic and rich in cysteine) is an extracellular protein associated with tissues exhibiting high rates of cell proliferation and matrix remodeling. The current work shows that the human melanoma cell lines IIB-MEL-LES, IIB-MEL-IAN, and IIB-MEL-J and different human metastatic melanomas expressed high levels of SPARC mRNA and protein. By western blot analysis we detected a single secreted 42-kDa band in human diploid fibroblasts-conditioned medium and a 45- to 40-kDa doublet in the three melanoma cell lines and all the metastatic melanomas tested. Part of the melanoma samples and cell lines showed an additional doublet of 36-34 kDa. SPARC mRNA was expressed by the three established cell lines, 14 metastatic melanoma samples, and tumors raised in nude mice, and no spliced variants were found. The heterogeneous pattern of SPARC secreted by human melanoma cells is the result of post-translational glycosylation and a specific extracellular leupeptin-inhibitable cleavage. Unlike human fibroblasts, melanoma cells did not overexpress SPARC on addition of TGF-beta. Immunohistochemical analysis showed that SPARC was strongly expressed in 100% of primary melanomas (7 of 7) and metastatic melanomas (29 of 29), moderately expressed in most of the positive dysplastic nevi (13 of 14), and only weakly expressed in nevocellular nevi (4 of 25). Normal melanocytes did not express SPARC. The data suggest that the expression of SPARC is associated with the neoplastic progression of human melanoma.
Assuntos
Melanoma/patologia , Osteonectina/biossíntese , Transformação Celular Neoplásica , Regulação da Expressão Gênica/efeitos dos fármacos , Glicosilação , Humanos , Imuno-Histoquímica , Linfotoxina-alfa/farmacologia , Melanoma/química , Melanoma/secundário , Metástase Neoplásica/genética , Osteonectina/genética , RNA Mensageiro/análise , Células Tumorais Cultivadas/químicaRESUMO
Acquisition of invasive/metastatic potential is a key event in tumor progression. Cell surface glycoproteins and their respective matrix ligands have been implicated in this process. Recent evidence reveals that the secreted glycoprotein SPARC (secreted protein, acidic and rich in cysteine) is highly expressed in different malignant tissues. The present study reports that the suppression of SPARC expression by human melanoma cells using a SPARC antisense expression vector results in a significant decrease in the in vitro adhesive and invasive capacities of tumor cells, completely abolishing their in vivo tumorigenicity. This is the first evidence that SPARC plays a key role in human melanoma invasive-metastatic phenotype development.
Assuntos
Movimento Celular/genética , Regulação Neoplásica da Expressão Gênica , Melanoma Experimental/patologia , Melanoma/patologia , Oligonucleotídeos Antissenso/genética , Osteonectina/genética , Animais , Adesão Celular/genética , Divisão Celular/genética , Regulação para Baixo , Humanos , Melanoma/genética , Melanoma Experimental/genética , Camundongos , Transfecção , Células Tumorais CultivadasRESUMO
Cytokine gene transfer to tumor cells has been demonstrated to induce tumor rejection in different murine models. However, controversial results were presented for different cytokines. In order to study the antitumorigenic activity that has been proposed for IL-6, the poorly immunogenic melanoma B16 and the colon adenocarcinoma CT26-murine cell lines, were transduced with recombinant retrovirus expressing rat IL-6. In vivo studies showed that IL-6-producing-B 16 cells inoculated s.c. in syngeneic mice, exhibited reduced tumorigenicity compared to vector-transduced B 16 cells. The histology of growing IL-6-producing tumors showed a "pseudo-nodular" pattern which correlated with a strong inhibition of the in vitro invasive capacity of these cells. IL-6-producing-B 16 cells did not develop tumors in athymic nude mice suggesting that the antitumor effect is not mediated by a normal host-T- and B-cell response. In contrast, IL-6-producing CT26 cells grew as tumors in syngeneic mice with a faster growth rate than parental and vector-transduced cells, in accordance with an increased in vitro growth kinetics. These results indicate that IL-6 expression by tumor cells demonstrate different effects depending on the tumor cell model.
Assuntos
Interleucina-6/genética , Células Tumorais Cultivadas/imunologia , Adenocarcinoma/genética , Adenocarcinoma/imunologia , Adenocarcinoma/patologia , Animais , Linfócitos B/imunologia , Divisão Celular , Neoplasias do Colo/genética , Neoplasias do Colo/imunologia , Neoplasias do Colo/patologia , Modelos Animais de Doenças , Expressão Gênica , Engenharia Genética , Cinética , Melanoma Experimental/genética , Melanoma Experimental/imunologia , Melanoma Experimental/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Nus , Invasividade Neoplásica/genética , Invasividade Neoplásica/imunologia , Invasividade Neoplásica/patologia , Transplante de Neoplasias , Ratos , Linfócitos T/imunologia , Transdução Genética , Transplante Isogênico , Células Tumorais Cultivadas/patologiaRESUMO
Previous studies from our laboratory have demonstrated that human melanoma cell lines and tumors expressed high levels of the extracellular protein SPARC. In order to demonstrate its role in human melanoma progression, IIB-MEL-LES human melanoma cells were transfected with SPARC full length c-DNA in the antisense orientation. In vivo studies demonstrated that all the control mice injected with parental cells developed tumors, while none of the mice injected with cells obtained from three different clones with diminished levels of SPARC expression, developed tumors. These studies suggest that SPARC may play a key role in human melanoma progression.
Assuntos
Melanoma/patologia , Osteonectina/fisiologia , Animais , Northern Blotting , Western Blotting , Células Clonais , DNA Antissenso/genética , Humanos , Masculino , Melanoma/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Osteonectina/metabolismo , Fatores de Tempo , Células Tumorais CultivadasRESUMO
Two human melanoma cell lines, derived from metastases of two patients with epithelioid malignant amelanotic melanomas, and designated IIB-MEL-LES and IIB-MEL-IAN, have been established. Both cell lines have been in continuous culture over 2 years and were propagated continuously for 85 and 75 serial passages, respectively. Morphologically, IIB-MEL-LES is composed predominantly of spindle shaped cells, whereas IIB-MEL-IAN grows as a monolayer of cuboid and stellate shaped cells with many rounded cells in suspension. Immunocytochemical studies revealed that both cell lines express S-100 protein, vimentin, and GD3 and GD2 gangliosides but are negative for keratin and collagen. Both cell lines express HLA class I and HLA-DR antigens in variable proportions. The MAGE-1 gene is expressed only by the IIB-MEL-IAN cell line, as revealed by PCR analysis. Cytogenetic analysis of both cell lines revealed abnormal karyotypes; the modal chromosome numbers of IIB-MEL-LES and IIB-MEL-IAN were 48 and 81, respectively. IIB-MEL-LES cells presented rearrangements in chromosomes 1, 14 and X, gains in chromosomes 10, 20, and 21 losses in chromosomes 15 and Y. The most frequent markers observed in IIB-MEL-IAN cells were 7q+, 10p+, 2p+, i(6p), 2q+, and 10q-. Clonal gains were observed in chromosomes 12 and 21, whereas losses were seen in chromosomes 1, 2, 3, 4, 6, 7, 11, and 17. Both cell lines were capable of forming colonies in soft agar and developed tumors when transplanted into nude mice, reproducing and maintaining the characteristics of the original tumors. These cell lines and their xenografts appear to provide useful systems for studying the biology, genetics and histogenesis of human malignant melanoma and could be utilized for the development of melanoma vaccines.
Assuntos
Imuno-Histoquímica , Melanoma Amelanótico/genética , Melanoma Amelanótico/patologia , Células Tumorais Cultivadas , Adulto , Animais , Sequência de Bases , Enzimas de Restrição do DNA , DNA de Neoplasias/análise , DNA de Neoplasias/química , Gangliosídeos/análise , Antígenos HLA-DR/análise , Antígenos de Histocompatibilidade Classe I/análise , Humanos , Imunofenotipagem , Cariotipagem , Masculino , Camundongos , Camundongos Nus , Dados de Sequência Molecular , Metástase Neoplásica , Transplante de Neoplasias , Proteínas S100/análise , Vimentina/análiseRESUMO
High levels of cytosolic cathepsin D expression have been associated with poor prognosis in breast cancer node-negative patients. In this work, we provide evidence that three cell lines established from human metastatic melanomas--IIB-MEL-J, IIB-MEL-LES, and IIB-MEL-IAN--express high levels of procathepsin D mRNA. IIB-MEL-J cells secreted into the conditioned media about 30% of the newly synthesized protein, which was active at acidic pH. Melanoma tumors arising in nude mice after injection of the three different cell lines expressed high levels of procathepsin D mRNA. Moreover, 13 human metastatic melanomas expressed variable levels of procathepsin D mRNA. To study the possible association between cathepsin D expression and melanoma development, samples corresponding to 10 primary tumors, 11 metastatic melanomas, 10 dysplastic nevi, 27 nevocellular nevi, and normal melanocytes were studied by immunohistochemistry for cathepsin D-specific staining. We found that cathepsin D was expressed in all of the dysplastic nevi and primary and metastatic melanomas tested but in only 18% of nevocellular nevi (five of 27), whereas normal melanocytes showed no cathepsin D expression. The overall data indicate that cathepsin D is expressed at a high level by melanoma cells, and because of its expression in preneoplastic lesions, it may be associated with melanoma development.
Assuntos
Catepsina D/análise , Síndrome do Nevo Displásico/metabolismo , Melanoma/secundário , Catepsina D/genética , Catepsina D/metabolismo , Meios de Cultivo Condicionados , Expressão Gênica , Humanos , Imuno-Histoquímica , Melanoma/química , Melanoma/genética , RNA Mensageiro/análise , Células Tumorais CultivadasRESUMO
FC-2.15 is an IgM monoclonal antibody (MAb) obtained by immunizing Balb/c mice with tumor epithelial cells from a human undifferentiated primary breast carcinoma. FC-2.15 reacts with 93.9% (31/33) of human breast primary tumors, independently of their histology and hormone receptor content. Moreover, FC-2.15 reacts with 79.6 +/- 13.8% (mean +/- SD) of total breast malignant tumor cells and with 88.7 +/- 9.9% of proliferating tumor cells. It recognizes other neoplasia such as colon cancer, squamous carcinoma and melanoma. Among the normal tissues examined, strong cross-reactivity was found with kidney proximal convolute tubules, bone marrow myeloid progeny, peripheral granulocytes and large bowel epithelium. Through Western blots, FC-2.15 recognizes three major bands of Mr 160 kDa, 130 kDa and 115 kDa in membrane extracts of MCF-7 cells grown in nude mice and of human breast carcinoma and three major bands of 250 kDa, 185 kDa and 105 kDa in membrane extracts of peripheral granulocytes. This MAb mediates complement- cytotoxicity against malignant cells, reducing the clonogenic capacity of breast primary tumor cells and MCF-7 cells to 35.6 +/- 41.2% and 11.7 +/- 4.8% of control values respectively, whereas that of normal bone marrow cells is not affected (104.7 +/- 17.4%).
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Antineoplásicos/imunologia , Neoplasias da Mama/imunologia , Neoplasias/imunologia , Animais , Anticorpos Monoclonais/isolamento & purificação , Anticorpos Monoclonais/uso terapêutico , Anticorpos Antineoplásicos/isolamento & purificação , Especificidade de Anticorpos , Antígenos de Neoplasias/química , Antígenos de Neoplasias/imunologia , Biomarcadores Tumorais/imunologia , Purging da Medula Óssea , Neoplasias da Mama/terapia , Citotoxicidade Imunológica , Feminino , Humanos , Imunização , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Peso Molecular , Células Tumorais Cultivadas/imunologia , Ensaio Tumoral de Célula-TroncoRESUMO
The MCF-7 breast carcinoma cell line can be separated by Percoll density gradient centrifugation into several subpopulations, A to F, one of which (E) has been previously suggested to be highly enriched in stem cells. The anchorage-independent growth of the different fractions and its sensitivity to estradiol (E2) and tamoxifen (TAM) was assayed. The anchorage-independent growth capacity of the different fractions was E greater than A greater than B greater than D greater than C,F. The E fraction had the highest clonogenic index (6.62 +/- 1.18) and was unaffected by E2 or TAM. The karyotypic analysis of the E fraction revealed features similar to those of the unfractionated cell line. It is suggested that the high growth rate of fraction E is due to an enrichment in stem cells and not to the existence of a different clone.
Assuntos
Neoplasias da Mama/patologia , Estradiol/farmacologia , Células-Tronco Neoplásicas/efeitos dos fármacos , Tamoxifeno/farmacologia , Neoplasias da Mama/genética , Adesão Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Marcadores Genéticos , Humanos , Cariotipagem , Células-Tronco Neoplásicas/citologiaRESUMO
In human breast cancer the proliferating cells appear to differ from those containing estrogen receptors (ER) as shown by studies on isolated cellular subpopulations. In this paper the in vitro effect of 17-beta-estradiol on cell proliferation in 30 primary breast tumors was studied. The effect of several estradiol concentrations was assayed, and the influence of diethylstilbestrol, tamoxifen, and nafoxidine was also tested. The response to these compounds was measured through the thymidine labeling index (TLI). When exposed to 10(-9) mol/l and 10(-8) mol/l estradiol, 14 of 19 ER-positive tumors and six of 11 ER-negative tumors were induced to further proliferate. The TLI increase over the control was 219% (P less than 0.05) at 10(-9) mol/l E2 and 258% (P less than 0.05) at 10(-8) mol/l E2 for ER-positive tumors, and 233% (0.1 less than P less than 0.2) at 10(-9) mol/l E2 and 321% (0.1 less than P less than 0.2) at 10(-8) mol/l E2 for ER-negative tumors. The addition of diethylstilbestrol and antiestrogens in vitro inhibited, to varying degrees, the estradiol-induced increase in the TLI irrespective of the ER-status. The response to E2 was correlated with the expression of the ras p21 protein and carcinoembryonic antigen. It was found that the ras p21 protein is preferentially expressed in ER-negative tumors, the opposite being true for carcinoembryonic antigen. The ras p21 protein is preferentially expressed in those ER-positive tumors that do not respond to estradiol with an increase in the TLI.
Assuntos
Neoplasias da Mama/patologia , Estradiol/farmacologia , Neoplasias da Mama/análise , Antígeno Carcinoembrionário/análise , Divisão Celular/efeitos dos fármacos , Replicação do DNA/efeitos dos fármacos , DNA de Neoplasias/análise , Dietilestilbestrol/farmacologia , Humanos , Nafoxidina/farmacologia , Proteínas de Neoplasias/análise , Neoplasias Hormônio-Dependentes/análise , Neoplasias Hormônio-Dependentes/patologia , Proteínas Proto-Oncogênicas/análise , Proteínas Proto-Oncogênicas p21(ras) , Receptores de Estrogênio/análise , Estimulação Química , Tamoxifeno/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
Exponentially growing MCF7 human breast cancer cells were separated in Percoll gradients into six different fractions of increasing density (A to F). These fractions could be subcultured and were found to contain different cellular subpopulations as defined by the following criteria: ability to generate other cellular subpopulations; growth rate; DNA synthesis; and expression of estrogen receptors, ras oncogene-encoded protein p21, and carcinoembryonic antigen. One of the minor fractions (E), which contained about 5% of the total cell number, appeared to contain the stem cells, on the basis of the following criteria: (i) its ability to reproduce the other cellular subpopulations, (ii) its high rate of growth and DNA synthesis, and (iii) the inability of the other subpopulations to generate it. The most differentiated subpopulation appeared to be the densest one (F), since it was the slowest growing and appeared to be the end point of the other subpopulations.
Assuntos
Neoplasias da Mama/patologia , Separação Celular/métodos , Modelos Biológicos , Neoplasias Hormônio-Dependentes/patologia , Células Tumorais Cultivadas/classificação , Antígeno Carcinoembrionário/análise , Centrifugação com Gradiente de Concentração , Inibição de Contato , Replicação do DNA , Feminino , Humanos , Proteínas de Neoplasias/análise , Povidona , Receptores de Estrogênio/análise , Dióxido de Silício , Células Tumorais Cultivadas/análise , Células Tumorais Cultivadas/patologiaRESUMO
Es conocido que los osteoclastos son ricos en fosfatasa acida tartrato-resistente (FATR) y se ha sugerido que esta podria liberarse al torrente sanguineo durante el desarrollo de metastasis oseas. En el presente trabajo se han medido en 72 pacientes con cancer (38 con metastasis oseas y 34 sin ellas), los niveles de FATR y fosfatasa alcalina (FALC), la cual hasta el momento ha sido el marcador serico que mejor ha reflejado la presencia de metastasis oseas. Los limites superiores para los dadores normales fueron: 2.9 UI para la FATR y 50 UI para la FALC. La FATR estuvo elevada en el 76% de los pacientes con metastasis mientras que la FALC estuvo en el 66% de dichos casos. Ningun paciente sin metastasis oseas presento FATR elevada mientras que un 12% tuvo FALC por encima de los valores normales. El 89% de los pacientes metastasicos presentaron por lo menos una de las 2 enzimas elevadas.Todo esto demuestra que la FATR es un buen marcador de la presencia de metastasis oseas y que su dosaje deberia complementar al de la FALC
Assuntos
Humanos , Masculino , Feminino , Fosfatase Alcalina , Neoplasias Ósseas , TartaratosRESUMO
Es conocido que los osteoclastos son ricos en fosfatasa acida tartrato-resistente (FATR) y se ha sugerido que esta podria liberarse al torrente sanguineo durante el desarrollo de metastasis oseas. En el presente trabajo se han medido en 72 pacientes con cancer (38 con metastasis oseas y 34 sin ellas), los niveles de FATR y fosfatasa alcalina (FALC), la cual hasta el momento ha sido el marcador serico que mejor ha reflejado la presencia de metastasis oseas. Los limites superiores para los dadores normales fueron: 2.9 UI para la FATR y 50 UI para la FALC. La FATR estuvo elevada en el 76% de los pacientes con metastasis mientras que la FALC estuvo en el 66% de dichos casos. Ningun paciente sin metastasis oseas presento FATR elevada mientras que un 12% tuvo FALC por encima de los valores normales. El 89% de los pacientes metastasicos presentaron por lo menos una de las 2 enzimas elevadas.Todo esto demuestra que la FATR es un buen marcador de la presencia de metastasis oseas y que su dosaje deberia complementar al de la FALC