RESUMO
Cross-talk between hematopoietic stem cells (HSCs) and bone marrow stromal cells (BMSCs) is essential for HSCs regulation and leukemogenesis. Studying bone marrow of myelodysplasia patients, a pre-leukemic condition, we found mRNA overexpression of vascular endothelial growth factor A (VEGFA) in CD34+ HSCs and semaphorin 3A (SEMA3A) in BMSCs. To better understand the role of VEGFA and SEMA3A in leukemogenesis, we recruited 30 myelodysplastic syndrome (MDS) patients, 29 acute myeloid leukemia (6 secondary to MDS) patients and 12 controls. We found higher VEGFA expression in de novo AML patients (without prior MDS) group (p=0.0073) and higher SEMA3A expression in all BMSCs patient's samples compared to control group. We then overexpressed VEGFA in an acute myelogenous leukemia cell line, KG1 cells, and in normal CD34+ cells. This overexpression increased KG1 (p=0.045) and CD34+ cell (p=0.042) viability and KG1 (p=0.042) and CD34+ cell (p=0.047) proliferation. Moreover, KG1 and CD34+ cells overexpressing VEGFA also had increased proliferation when co-cultured with human marrow stromal HS5 cells (p=0.045 and p=0.02, respectively). However, co-culture of these transformed cells with HS5 cells overexpressing SEMA3A reduced KG1 (p=0.004) and CD34+ (p=0.009) proliferation. Co-culture of KG1 transformed cells with HS27 cells overexpressing SEMA3A reduced KG1 proliferation as well (p=0.01). To investigate whether the dominant SEMA3A effect over VEGFA could be due to competition for neuropilin1 receptor (NRP1), we performed immunoprecipitation with anti-NRP1 antibody of cell extracts of co-cultured KG1 and HS5 cells, induced or not by VEGFA and SEMA3A recombinant proteins. Results showed a preferential association of NRP1 with SEMA3A, suggesting that SEMA3A can partially reverse the effects caused by the VEGFA preventing its binding with the NRP1 receptor. Since both hematopoietic cells, leukemic and normal, showed similar behavior, we suppose that the attempt to reversion of VEGF effects by SEMA3A is a homeostatic phenomenon in the hematopoietic niche. Finally, we conclude that VEGFA overexpression confers AML cell advantages and SEMA3A may partially reverse this effect; thus, SEMA3A protein combined with VEGFA inhibitors could be beneficial for AML treatment.
Assuntos
Neuropilina-1/metabolismo , Semaforina-3A/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Células da Medula Óssea , Linhagem Celular Tumoral/metabolismo , Linhagem Celular Tumoral/patologia , Feminino , Humanos , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patologia , Masculino , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/patologia , Pessoa de Meia-Idade , Síndromes Mielodisplásicas/metabolismo , Síndromes Mielodisplásicas/patologia , Ligação Proteica , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Transfecção , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
A number of experimental and clinical reports suggest the involvement of oxidative stress in pathophysiology of epilepsy. Topiramate, a new antiepileptic drug, induces antioxidant effect in epileptic animals. However, to date, no further studies appear to be carried out in order to demonstrate the ability of topiramate to act as antioxidant. Therefore, the objective of this work was to evaluate the in vitro superoxide (O2(·-)), hydroxyl radical (OH·), hypochlorous acid (HOCl), hydrogen peroxide (H2O2), singlet oxygen ((1)O2) and peroxynitrite (ONOO(-)) scavenging capacity of topiramate in comparison with reference compounds. In addition, we investigated the possible antitumour activity of this compound in some cancer cell lines. Topiramate displays a scavenging capacity compared to the reference compound, with the exception of ONOO(-), although it was less efficient than nordihydroguaiaretic acid, dimethylthiourea, ascorbic acid, sodium pyruvate and glutathione for O2(·-), OH·, HOCl, H2O2 and (1)O2(P < 0.0001), respectively, and not induced significant growth inhibition in cancer cell lines. The direct antioxidant properties of topiramate could explain the neuroprotective effects attributed to this compound and suggest its use as chemopreventive agent in a future.
Assuntos
Antioxidantes/farmacologia , Sequestradores de Radicais Livres/metabolismo , Frutose/análogos & derivados , Linhagem Celular Tumoral/classificação , Linhagem Celular Tumoral/patologia , Relação Dose-Resposta a Droga , Frutose/farmacologia , Humanos , Oxirredução/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , TopiramatoRESUMO
El cáncer es una enfermedad multifactorial caracterizada por anormalidad en el crecimiento celular provocado por factores ambientales y múltiples cambios en la expresión de los genes. El cáncer de mama representa una de las enfermedades que más afecta a la población femenina mundial. Actualmente, se ha mostrado un auge en las investigaciones sobre compuestos extraídos de diversas plantas y origen sintético con posibles agentes antitumorales, entre los que se pueden nombrar los derivados sintéticos de quinolinas. Realizado estudio del grupo sintético indeno (2,1-c) quinolinas y sus efectos citotóxicos sobre líneas de cáncer de mama SKBr3 yMCF-7. La evaluación de la actividad citotóxica de estos compuestos fue determinada a través del método del MTT, se calcularon los valores de concentración inhibitoria, los índices de selectividad y combinaciones con taxanos. Los resultados obtenidos muestran una alta selectividad de los compuestos sintéticos y natural evaluados hacia las líneas celulares MCF-7 y SKBr3 con respecto a las células control (fibroblastos dérmicos humanos); además de una interacción de tipo potenciación en la combinación de un derivado quinolínico y la drogataxano. Con todo esto se puede inferir que solo ciertos compuestos que tuvieron modificaciones químicas realizadas sobre el compuesto patrón del grupo sintético indeno (2,1-c) quinolinas mostraron una inhibición sobre la viabilidad celular sobre las líneas tumorales SKBr3 y MCF-7, conforme aumenta la dosis de los compuestos, además de evidenciar la efectividad de compuestos derivados de quinolinas como moléculas con características antineoplásicas
The cancer is a multifactorial disease characterized by abnormal cell growth caused by environmental factors and for multiple changes in the gene expression. The breast cancer is a disease that affects the female population in worldwide. Currently, there is shown an increment in the research on compounds that we extracted from various plants and synthetic origin with potential antitumor agents, among which can be namedthe synthetic derivatives of the quinolines. A study was performed on an group of synthetic indeno (2,1-c) quinolines and their cytotoxic effects on breast cancer cell lines SKBR3 and MCF-7 was performed. The evaluation of the cytotoxic activity of these compounds was determined through the MTT method; we were calculated inhibitory concentration values, the rates of selectivity and the drug combinations with taxanes. The results show a high selectivity of the natural and the synthetic compounds evaluated towards the cell lines MCF-7 and SKBR3 cells with respect to control (dermal fibroblasts human), besides an enhancement type interaction in the combination of a drug derivative and the quinolinic taxanes. With all this we can infer that only certain compounds had chemical modifications performed on the composite pattern of synthetic group indeno (2, 1-c) quinolines showed an inhibition on cell viability on tumor cell lines SKBR3 and MCF-7, with increasing dose of the compounds, also showing the effectiveness of compounds derived from the quinolines as molecules with the antineoplastic properties and his characteristics
Assuntos
Feminino , Citotoxicidade Celular Dependente de Anticorpos , Neoplasias da Mama/prevenção & controle , Neoplasias da Mama/terapia , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Tratamento Farmacológico/métodos , Quinolinas/uso terapêutico , Vacinas Sintéticas/uso terapêutico , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Linhagem Celular Tumoral , Linhagem Celular Tumoral/patologia , OncologiaRESUMO
BACKGROUND: Cancer-initiating cells (CICs) are proposed to be responsible for the generation of metastasis and resistance to therapy. Accumulating evidences indicates CICs are found among different human cancers and cell lines derived from them. Few studies address the characteristics of CICs in cervical cancer. We identify biological features of CICs from four of the best-know human cell lines from uterine cervix tumors. (HeLa, SiHa, Ca Ski, C-4 I). METHODS: Cells were cultured as spheres under stem-cell conditions. Flow cytometry was used to detect expression of CD34, CD49f and CD133 antigens and Hoechst 33342 staining to identify side population (SP). Magnetic and fluorescence-activated cell sorting was applied to enrich and purify populations used to evaluate tumorigenicity in nude mice. cDNA microarray analysis and in vitro radioresistance assay were carried out under standard conditions. RESULTS: CICs, enriched as spheroids, were capable to generate reproducible tumor phenotypes in nu-nu mice and serial propagation. Injection of 1 × 10³ dissociated spheroid cells induced tumors in the majority of animals, whereas injection of 1 × 105 monolayer cells remained nontumorigenic. Sphere-derived CICs expressed CD49f surface marker. Gene profiling analysis of HeLa and SiHa spheroid cells showed up-regulation of CICs markers characteristic of the female reproductive system. Importantly, epithelial to mesenchymal (EMT) transition-associated markers were found highly expressed in spheroid cells. More importantly, gene expression analysis indicated that genes required for radioresistance were also up-regulated, including components of the double-strand break (DSB) DNA repair machinery and the metabolism of reactive oxygen species (ROS). Dose-dependent radiation assay indicated indeed that CICs-enriched populations exhibit an increased resistance to ionizing radiation (IR). CONCLUSIONS: We characterized a self-renewing subpopulation of CICs found among four well known human cancer-derived cell lines (HeLa, SiHa, Ca Ski and C-4 I) and found that they express characteristic markers of stem cell, EMT and radioresistance. The fact that CICs demonstrated a higher degree of resistance to radiation than differentiated cells suggests that specific detection and targeting of CICs could be highly valuable for the therapy of tumors from the uterine cervix.
Assuntos
Células-Tronco Neoplásicas/efeitos da radiação , Neoplasias do Colo do Útero/patologia , Animais , Antígenos CD/metabolismo , Benzimidazóis/metabolismo , Biomarcadores Tumorais/metabolismo , Linhagem Celular Tumoral/patologia , Transformação Celular Neoplásica , Transição Epitelial-Mesenquimal/genética , Transição Epitelial-Mesenquimal/fisiologia , Feminino , Citometria de Fluxo , Células HeLa , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Análise em Microsséries , Células-Tronco Neoplásicas/metabolismo , Tolerância a Radiação , Esferoides Celulares/patologia , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/radioterapia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Small RNAs belong to a newly discovered strain of molecules. These molecules are composed of double strand RNA comprised by just about 19-31 nucleotides. They have two main characteristics that make them unique. Firstly, they are noncoding for proteins and second they interfere post-transcriptional with mRNA. This interfering action is the distinguishing hallmark, therefore known as interfering RNA or RNAi. There are three main subclasses of which micro-RNA and siRNA are the most widely studied. Interference RNAs participate in a myriad of cellular functions mainly through modulation of genetic expression. Due to these capabilities it has been used as therapeutic weapon in a number of diseases including cancer. It is known that both miRNA and siRNA participate in carcinogenesis, either inhibiting suppressor genes, or stimulating oncogenes. It has been demonstrated that manipulating small interfering RNAs in cell lines and animal models, the malignant and metastatic phenotype can be reversed. Up to now a few clinical trials using RNAi as a therapeutic agent have demonstrated some success and feasibility. It is forseeable that in the near future cancer treatment with small RNAs will be widely applicable, once the many constrains for its systemic application are surpassed.
Assuntos
Regulação Neoplásica da Expressão Gênica/genética , Terapia Genética/métodos , MicroRNAs/genética , Neoplasias/genética , Interferência de RNA , RNA Mensageiro/genética , RNA Interferente Pequeno/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/patologia , Transformação Celular Neoplásica/genética , Ensaios Clínicos como Assunto , Sistemas de Liberação de Medicamentos , Ensaios de Seleção de Medicamentos Antitumorais , Estudos de Viabilidade , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Genes Supressores de Tumor , Vetores Genéticos/uso terapêutico , Humanos , Masculino , MicroRNAs/biossíntese , Neoplasias/tratamento farmacológico , Neoplasias/terapia , Neoplasias/virologia , Oncogenes , RNA Mensageiro/antagonistas & inibidores , RNA Neoplásico/antagonistas & inibidores , RNA Neoplásico/biossíntese , RNA Neoplásico/genética , RNA Interferente Pequeno/genéticaRESUMO
Mast cell (MC) activation in the hamster cheek pouch cancerization model is associated with the increase in tumor cell proliferation, mediated in turn by tryptase, a protease released from mast cell granules after activation. Tryptase induces tumor cell proliferation through the activation of PAR-2 (protease activated receptor-2) on the plasma membrane of carcinoma cells. The therapeutic success of boron neutron capture therapy mediated by boronophenylalanine (BPA-BNCT) in tumor control in the hamster cheek pouch oral cancer model has been previously reported by our laboratory. Early effects of BPA-BNCT on tumors of the hamster cheek pouch include a reduction in DNA-synthesis with the concomitant decrease in the proliferation of malignant cells. The aim of the present study was to investigate the early histological changes in mast cells after BPA-BNCT in tumors and premalignant tissue of the hamster cheek pouch. Tumor-bearing pouches were treated with BPA-BNCT or beam only (neutron irradiation without prior administration of the boron compound) and sacrificed 1day after treatment. The samples were fixed in Carnoy fixative and stained with alcian blue-safranin to identify all the populations of mast cells. Total, active and inactive mast cells (MC) were counted in the connective tissue and the adventitious tissue underlying the pouch wall and at the base of the tumors in pouches treated with BPA-BNCT, in keeping with a previously described technique. BPA-BNCT induced a marked reduction in the total number of mast cells in the pouch (p<0.05). This reduction in the total number of mast cells was due to a reduction in mast cells at the base of the tumor (p<0.005) and it occurred at the expense of the active mast cells (p<0.05). A slight reduction that did not reach statistical significance also occurred in the amount of mast cells in the pouch wall (that corresponds to the premalignant tissue in tumor-bearing pouches), and in the adventitious tissue. In this case the reduction was seen in the inactive population. Both BPA-BNCT and beam only elicited a qualitative change in the secretion modality of the granule content. Although further studies are needed to evaluate the subcellular effect of BNCT on mast cell granule secretion, the reduction in cell proliferation induced by BPA-BNCT would be partially due to the decrease in total mast cells in the hamster check pouch.
Assuntos
Compostos de Boro/uso terapêutico , Linhagem Celular Tumoral/patologia , Mastócitos/patologia , Neoplasias Bucais/patologia , Fenilalanina/análogos & derivados , Lesões Pré-Cancerosas/patologia , Radiossensibilizantes/uso terapêutico , Animais , Terapia por Captura de Nêutron de Boro/métodos , Contagem de Células , Proliferação de Células , Bochecha/patologia , Bochecha/efeitos da radiação , Cricetinae , Mastócitos/efeitos da radiação , Neoplasias Bucais/radioterapia , Fenilalanina/uso terapêutico , Lesões Pré-Cancerosas/radioterapiaRESUMO
BACKGROUND AND OBJECTIVE: Circulating tumor cells have been shown to correlate positively with metastatic disease state in patients with advanced cancer. We have demonstrated the ability to detect melanoma cells in a flow system by generating and detecting photoacoustic waves in melanoma cells. This method is similar to flow cytometry, although using photoacoustics rather than fluorescence. Previously, we used piezoelectric films as our acoustic sensors. However, such films have indicated false-positive signals due to unwanted direct interactions between photons from the high laser fluence in the flow system and the film itself. We have adapted an optical detection scheme that obviates the need for piezoelectric films. STUDY DESIGN/MATERIALS AND METHODS: Our photoacoustic system comprised a tunable laser system with an output of 410-710 nm with a pulse duration of 5 nanoseconds. The light was delivered by optical fiber to a glass microcuvette that contained saline buffer suspensions of melanoma and white blood cells. We used a continuous HeNe laser to provide a probe beam that reflected off of a glass and water interface in close proximity to the microcuvette. The beam was detected by a high-speed photodiode. When a photoacoustic wave was generated in the microcuvette, the wave propagated and changed the reflectance of the beam due to index of refraction change in the water. This perturbation was used to detect the presence of melanoma cells. RESULTS: We determined a detection threshold of about one individual melanoma cell with no pyroelectric noise indicated in the signals. CONCLUSIONS: The optical reflectance method provides sensitivity to detect small numbers of melanoma cells without created false-positive signals from pyroelectric interference, showing promise as a means to perform tests for circulating melanoma cells in blood samples.
Assuntos
Acústica/instrumentação , Separação Celular/instrumentação , Tecnologia de Fibra Óptica/instrumentação , Citometria de Fluxo/instrumentação , Lasers Semicondutores , Células Neoplásicas Circulantes/patologia , Linhagem Celular Tumoral/patologia , Separação Celular/métodos , Citometria de Fluxo/métodos , Humanos , Aumento da Imagem/instrumentação , Técnicas In Vitro , Melaninas/metabolismo , Melanoma/patologia , Reconhecimento Automatizado de Padrão , Probabilidade , Neoplasias Cutâneas/patologiaRESUMO
In recent years, much attention has been focused on polysaccharides isolated from natural sources. The aim of this study was to investigate the in vitro and in vivo antitumor properties of a sulfated polysaccharide isolated from the seaweed C. feldmannii (Cf-PLS). Hematological, biochemical and histopathological analyses were performed in order to evaluate the toxicological aspects related to Cf-PLS treatment. Its effects on the immunological system were also investigated. The Cf-PLS did not show any significant in vitro cytotoxicity at the experimental exposure levels that were used, but showed in vivo antitumor effect. The inhibition rates of sarcoma 180 tumor development were 48.62 and 48.16% at the doses of 10 and 25 mg kg(-1), respectively. In addition, Cf-PLS was also able to increase the response elicited by 5-fluorouracil (5-FU) from 48.66 to 68.32%. The histopathological analysis of liver and kidney showed that both organs were moderately affected by Cf-PLS-treatment. Neither enzymatic activity of alanine aminotransferase nor urea or creatinine levels were significantly altered. In hematological analysis, leucopeny was observed after 5-FU treatment, but this effect was prevented when the treatment was associated with the Cf-PLS. It was also demonstrated that Cf-PLS acts as an immunomodulatory agent, raising the production of specific antibodies, and increasing the production of OVA-specific antibodies. It also induced a discreet hyperplasia of lymphoid folicules of the white pulp in the spleen of treated mice. In conclusion, Cf-PLS has some interesting anticancer activity that could be associated with its immunostimulating properties.
Assuntos
Antineoplásicos/farmacologia , Linhagem Celular Tumoral/efeitos dos fármacos , Polissacarídeos/farmacologia , Rodófitas/química , Sarcoma 180/tratamento farmacológico , Animais , Formação de Anticorpos/efeitos dos fármacos , Antineoplásicos/toxicidade , Linhagem Celular Tumoral/patologia , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Sinergismo Farmacológico , Quimioterapia Combinada , Feminino , Fluoruracila/farmacologia , Humanos , Fatores Imunológicos/farmacologia , Fatores Imunológicos/toxicidade , Camundongos , Extratos Vegetais/farmacologia , Extratos Vegetais/toxicidade , Polissacarídeos/toxicidade , Sarcoma 180/patologia , Alga Marinha , Sulfatos/farmacologia , Sulfatos/toxicidade , Testes de ToxicidadeRESUMO
OBJECTIVE AND DESIGN: Gross cystic disease (GCD) of the breast is reported to occur in 7% of women in the developed world and, although not premalignant, is thought to be associated with an increased risk of breast cancer. Hormone and growth factor concentration levels were measured in breast cyst fluid (BCF) to correlate them with their mitogenic activity in tumour (MCF-7) or nontransformed (MCF-10A) cells. RESULTS: Oestradiol (E2), oestrone (E1), E2-sulfate (E2-S), E1-sulfate (E1-S) and epidermal growth factor (EGF) concentrations were, as expected, significantly higher in type I than in type II cysts, while transforming growth factor-beta 2 (TGF-beta2) showed higher levels in type II cysts. Fifty per cent of the BCF samples stimulated [3H]-thymidine incorporation into MCF-7 cells while 34.5% inhibited this parameter. In MCF-10A cells, most BCF samples were stimulatory (85%). E2, E1 and EGF concentrations in BCF samples correlated significantly and positively with cell proliferation in MCF-7 cells, whereas a significant negative correlation was found for TGF-beta2. In MCF-10A cells, only E2-S and E1-S exhibited significant positive correlation, whereas a significant negative correlation was found for TGF-beta2. Progesterone (Pg), E2 and EGF incubated under the same conditions had a stimulatory effect on [3H]-thymidine incorporation into MCF-7 cells, whereas TGF-beta2 inhibited this parameter. Pg, E2, E1 and EGF significantly stimulated this parameter in MCF-10A cells. CONCLUSIONS: The stimulatory action of BCF on cell proliferation in a model of human breast epithelial cells could partly explain the increased incidence of breast cancer in cyst-bearing women.
Assuntos
Cisto Mamário/metabolismo , Linhagem Celular Tumoral/patologia , Hormônio do Crescimento/metabolismo , Hormônios/metabolismo , Adulto , Líquidos Corporais/química , Líquidos Corporais/metabolismo , Linhagem Celular Tumoral/metabolismo , Proliferação de Células , Fator de Crescimento Epidérmico/análise , Células Epiteliais/patologia , Feminino , Humanos , Concentração de Íons de Hidrogênio , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/análise , Fator de Crescimento Insulin-Like I/análise , Potássio/análise , Sódio/análise , Fator de Crescimento Transformador beta/análiseRESUMO
Adenoid cystic carcinoma (ACC) is a frequent malignant salivary gland neoplasm presenting different growth patterns described as tubular, cribriform and solid, which represent distinct differentiation stages. Cell lines originated from ACCs grown inside three-dimensional environments have not been capable to reproduce all in vivo ACC growth patterns. As ACC cells in vivo present replicated basement membrane, to mimic this situation in vitro ACC cells (CAC2 cells) were grown on the top of a reconstituted basement membrane (Matrigel). Phenotype differences were assessed by light, fluorescence and transmission electron microscopy. The cultures grown on the top of Matrigel presented three-dimensional arrangement of cells intercepted by cellular cords. At these, cell nests pseudocyst formations were observed. This morphological structure entirely reproduced the cribriform growth pattern of ACC. We suggest that the cribriform differentiation of ACC in culture is dependent of proteins and growth factors associated in a bi-dimensional structure.
Assuntos
Materiais Biocompatíveis/farmacologia , Carcinoma Adenoide Cístico/patologia , Colágeno/farmacologia , Laminina/farmacologia , Proteoglicanas/farmacologia , Neoplasias das Glândulas Salivares/patologia , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/patologia , Combinação de Medicamentos , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/ultraestrutura , Humanos , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Microscopia de Contraste de FaseRESUMO
BACKGROUND/AIM: Pro-inflammatory cytokines and chemokines, such as interleukin (IL) 8, are important mediators of hepatic injury and repair following an insult. The purpose of this work was to study the regulation of IL-8 by IL-10 and IL-4 in HepG2 cells treated with acetaldehyde (Ac). METHODS: HepG2 cells were pretreated with IL-10 or IL-4 before exposure to Ac, examining IL-8 expression by reverse transcription polymerase chain reaction and Western blot. RESULTS: Ac treatment produced an increment in IL-8 induction and secretion that was prevented by IL-4 pretreatment, while IL-10 pretreatment failed to decrease Ac-induced IL-8 production. Consistent with these findings Ac increased NF-kappa B and AP-1 activation that were prevented by IL-4 but not by IL-10, findings accompanied by greater I kappa B-alpha levels in IL-4 but not IL-10 pretreated cells. In contrast to the pro-inflammatory role of IL-10 in HepG2, IL-10 did not show any change in the activation of NF-kappa B by Ac in WRL-68 cells, a human fetal hepatic cell line. Moreover, IL-10 did not induce the degradation of I kappa B-alpha in cellular extract from rat primary cultured cells. CONCLUSIONS: While the present findings demonstrate the anti-inflammatory role of IL-4 in preventing the expression of IL-8 by Ac, the regulation of chemokines by anti-inflammatory cytokines is complex and depends on the cellular lineage.
Assuntos
Acetaldeído/farmacologia , Carcinoma Hepatocelular/tratamento farmacológico , Interleucina-10/farmacologia , Interleucina-4/farmacologia , Interleucina-8/metabolismo , Neoplasias Hepáticas/tratamento farmacológico , Animais , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/patologia , Combinação de Medicamentos , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Proteínas I-kappa B/metabolismo , Interleucina-8/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Masculino , Inibidor de NF-kappaB alfa , NF-kappa B/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição AP-1/metabolismoRESUMO
Gastric cancer is the second most frequent type of neoplasia and also the second most important cause of death in the world. Virtually all the established cell lines of gastric neoplasia were developed in Asian countries, and western countries have contributed very little to this area. In the present study we describe the establishment of the cell line ACP01 and characterize it cytogenetically by means of in vitro immortalization. Cells were transformed from an intestinal-type gastric adenocarcinoma (T4N2M0) originating from a 48-year-old male patient. This is the first gastric adenocarcinoma cell line established in Brazil. The most powerful application of the cell line ACP01 is in the assessment of cytotoxicity. Solid tumor cell lines from different origins have been treated with several conventional and investigational anticancer drugs. The ACP01 cell line is triploid, grows as a single, non-organized layer, similar to fibroblasts, with focus formation, heterogeneous division, and a cell cycle of approximately 40 h. Chromosome 8 trisomy, present in 60 percent of the cells, was the most frequent cytogenetic alteration. These data lead us to propose a multifactorial triggering of gastric cancer which evolves over multiple stages involving progressive genetic changes and clonal expansion.
Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Adenocarcinoma/genética , Linhagem Celular Tumoral , Análise Citogenética/métodos , Neoplasias Gástricas/genética , Adenocarcinoma/patologia , Células Clonais , Criopreservação , Linhagem Celular Tumoral/patologia , Cariotipagem , Neoplasias Gástricas/patologia , Trissomia/genética , Trissomia/patologiaRESUMO
Gastric cancer is the second most frequent type of neoplasia and also the second most important cause of death in the world. Virtually all the established cell lines of gastric neoplasia were developed in Asian countries, and western countries have contributed very little to this area. In the present study we describe the establishment of the cell line ACP01 and characterize it cytogenetically by means of in vitro immortalization. Cells were transformed from an intestinal-type gastric adenocarcinoma (T4N2M0) originating from a 48-year-old male patient. This is the first gastric adenocarcinoma cell line established in Brazil. The most powerful application of the cell line ACP01 is in the assessment of cytotoxicity. Solid tumor cell lines from different origins have been treated with several conventional and investigational anticancer drugs. The ACP01 cell line is triploid, grows as a single, non-organized layer, similar to fibroblasts, with focus formation, heterogeneous division, and a cell cycle of approximately 40 h. Chromosome 8 trisomy, present in 60% of the cells, was the most frequent cytogenetic alteration. These data lead us to propose a multifactorial triggering of gastric cancer which evolves over multiple stages involving progressive genetic changes and clonal expansion.
Assuntos
Adenocarcinoma/genética , Linhagem Celular Tumoral , Análise Citogenética/métodos , Neoplasias Gástricas/genética , Adenocarcinoma/patologia , Linhagem Celular Tumoral/patologia , Cromossomos Humanos Par 8 , Células Clonais , Criopreservação , Humanos , Cariotipagem , Masculino , Pessoa de Meia-Idade , Neoplasias Gástricas/patologia , Trissomia/genética , Trissomia/patologiaRESUMO
OBJECTIVE: Present study addresses the issue whether apoptosis and necrosis increases the antigenicity of proteins recognized by antinuclear antibodies. MATERIAL AND METHODS: HEp-2 cells were cultured in standard conditions; apoptosis was induced by camptothecin and necrosis by mercuric chloride. Protein antigenicity of cell extracts was tested onto nitrocellulose membranes and probed with positive or negative sera for antinuclear antibodies by a luminescent-dot-ELISA system. RESULTS: Apoptotic changes in HEp-2 cells appeared by 24 hours of camptothecin exposure, meanwhile the necrotic features become visible earlier. Luminescence was significantly superior in ANA positive sera than in ANA negative controls. Antinuclear antibody sera recognized better the antigens from the apoptotic and necrotic cells than controls without chemical treatments. CONCLUSIONS: Apoptosis and necrosis increase the ANA binding by better availability of intracellular antigens, or by disclosing cryptic epitopes.
Assuntos
Anticorpos Antinucleares/imunologia , Apoptose/imunologia , Autoantígenos/imunologia , Doenças Autoimunes/imunologia , Anticorpos Antinucleares/sangue , Reações Antígeno-Anticorpo , Antígenos de Neoplasias/imunologia , Apoptose/efeitos dos fármacos , Doenças Autoimunes/patologia , Camptotecina/farmacologia , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/imunologia , Linhagem Celular Tumoral/patologia , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Fragmentos Fab das Imunoglobulinas/imunologia , Marcação In Situ das Extremidades Cortadas , Neoplasias Laríngeas/imunologia , Neoplasias Laríngeas/patologia , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/imunologia , Cloreto de Mercúrio/farmacologia , Doença Mista do Tecido Conjuntivo/sangue , Doença Mista do Tecido Conjuntivo/imunologia , Necrose , Proteínas de Neoplasias/imunologia , Esclerodermia Difusa/sangue , Esclerodermia Difusa/imunologia , Síndrome de Sjogren/sangue , Síndrome de Sjogren/imunologiaRESUMO
Nitric oxide (NO)-derived from T lymphocytes in an autocrine fashion can modulate events in the cell. However, the exact role of NO on the control of lymphocyte growth is controversial since both stimulation and inhibition have been demonstrated. Nitric oxide synthase (NOS) activity in normal and tumor T lymphocyte proliferation was studied here. Resting normal T lymphocytes displayed low levels of NOS activity that were slightly increased upon mitogenic stimulation. In contrast, BW5147 T lymphoma cells displayed higher basal levels than normal T lymphocytes that were significantly augmented when induced to proliferate. This activity was slightly modified in the presence of the calcium chelator EGTA and was blocked by competitive and irreversible NOS inhibitors, as well as by selective blockers of iNOS. Furthermore, tumor but not normal cell proliferation was impaired by NOS and iNOS blockers, while a calcium blocker only affected normal cell growth. iNOS expression, both at the protein and at the mRNA levels, was demonstrated on growing BW5147 cells but not on arrested tumor or normal lymphocytes. The contribution of iNOS to sustained proliferation of tumor cells is discussed.