RESUMEN
This brief review focuses on the emerging role of matrix metalloproteinase 11 (MMP-11) in cancer progression. It has recently been shown that MMP-11 is induced in adipose tissue by cancer cells as they invade their surrounding environment. MMP-11 negatively regulates adipogenesis by reducing pre-adipocyte differentiation and reversing mature adipocyte differentiation. Adipocyte dedifferentiation in turn leads to the accumulation of nonmalignant peritumoral fibroblast-like cells, which favor cancer cell survival and tumor progression. This MMP-11-mediated bi-directional cross-talk between invading cancer cells and adjacent adipocytes/pre-adipocytes highlights the central role that MMP-11 plays during tumor desmoplasia and represents a molecular link between obesity and cancer.
Asunto(s)
Adipocitos/enzimología , Metaloproteinasa 11 de la Matriz/metabolismo , Neoplasias/enzimología , Neoplasias/patología , Animales , Comunicación Celular , Progresión de la Enfermedad , Humanos , Estadificación de NeoplasiasRESUMEN
Impedance sensors in thick film technology have been tested as a tool for electric cell-substrate impedance sensing. The screen printed Pt electrodes have a width of 250-400 microm. Electrodes and the surrounding ceramic chip substrate could be homogeneously grown with L-929 and Hela cells. The performance of a screen printed interdigitated electrode structure (IDES) was compared with that of thin film structures with the same layout geometry. The thick film impedance sensors allowed to correctly record the morphological response of confluent Hela cell layers to stimulation with histamine. A thick film conductivity sensor also revealed impedance values which were dependent on cell growth on the electrode surface, even at a very low frequency range of approximately 1 Hz.
Asunto(s)
Técnicas Biosensibles/métodos , Electrodos , Técnicas Analíticas Microfluídicas/métodos , Animales , Procesos de Crecimiento Celular/fisiología , Cerámica/química , Impedancia Eléctrica , Fibroblastos/citología , Vidrio , Células HeLa , Humanos , Ratones , Platino (Metal)/químicaRESUMEN
The intracellular protease calpain, abundant in endothelial cells (EC), is assumed to be inactive under physiological conditions but may account for Ca2+ -linked pathophysiological events. However, nonstimulated EC contained autolyzed, activated calpain. Adding 12-48 microM calpain inhibitor I (CI) or 0.5-1 microM of the novel, membrane-permeable conjugate of calpastatin peptide-penetratin (CPP) caused rapid rounding and retraction of cultured EC (phase contrast, capacitance) and translocation of Syk, Rac, and Rho to the membrane, signifying activation upon inhibition of calpain. Isolated hearts (guinea pig) perfused with 12 microM CI or 0.5 muM CPP developed coronary leak. We conclude that calpain is constitutively active in EC and regulates vascular permeability by governing cell-cell attachment.
Asunto(s)
Calpaína/metabolismo , Permeabilidad de la Membrana Celular/fisiología , Células Endoteliales/citología , Células Endoteliales/fisiología , Animales , Células Cultivadas , Cobayas , Humanos , Venas Umbilicales/citologíaRESUMEN
The initial invasive processes during cancer development remain largely unknown. Stromelysin-3/matrix metalloproteinase 11 (ST3/MMP11) is associated with tumor invasion and poor prognosis. We present novel evidence that adipocytes present at human breast tumor invasive front are induced by cancer cells to express ST3. Using mouse syngeneic model, light and electron microscopy showed that in ST3-deficient mice but not in wild-type mice, forced cancer cell-adipocyte interaction/crosstalk results in adipocyte membrane alteration, allowing cancer cell fat infiltration and death. Thus, adipocytes are involved in initial cancer cell survival into connective tissue, and this effect is ST3 mediated. This suggested that ST3 might play a role in adipocyte metabolism. Accordingly, ST3-deficient mice exhibited fat excess and increased mRNA levels of peroxisome proliferator-activated receptor gamma (PPARgamma) and adipocyte protein 2 (aP2) adipogenic markers, indicating that, in vivo, ST3 negatively regulates fat homeostasis. Moreover, ST3-deficient mouse embryonic fibroblasts exhibited a dramatic enhanced potential to differentiate into adipocytes associated with increased PPARgamma and aP2 expression, and recombinant ST3 treatment reverted their differentiation. Thus, in vitro, ST3 reduces adipocyte differentiation in an autocrine manner. High fibroblasts/adipocytes ratio is a stroma feature, and peritumoral fibroblast origin remains debated. Our results support the concept that invading cancer cells aberrantly restore the negative ST3 function on adipogenesis into proximal adipocytes/preadipocytes, leading to the accumulation/maintenance of a particular peritumoral fibroblast subpopulation. Accordingly, in human breast tumors, we observed that ST3-expressing peritumoral fibroblasts are distinct from alpha-smooth muscle actin-expressing myofibroblasts. This constitutes the first report of implication of a MMP in cancer cell-adipocyte interaction/crosstalk during early steps of connective tissue invasion.
Asunto(s)
Adipocitos/citología , Adipogénesis/fisiología , Neoplasias de la Mama/patología , Comunicación Celular/fisiología , Metaloendopeptidasas/fisiología , Adipocitos/metabolismo , Animales , Neoplasias de la Mama/metabolismo , Diferenciación Celular/fisiología , Neoplasias del Colon/metabolismo , Neoplasias del Colon/patología , Embrión de Mamíferos , Femenino , Fibroblastos/citología , Fibroblastos/metabolismo , Humanos , Masculino , Metaloproteinasa 11 de la Matriz , Metaloendopeptidasas/biosíntesis , Metaloendopeptidasas/deficiencia , Ratones , Ratones Endogámicos BALB C , Invasividad NeoplásicaRESUMEN
Chemotherapeutic drugs affect the metabolism of tumor cells regardless of the specific target of action. Basic parameters of cell metabolism are extrusion of acids into the microenvironment and oxygen consumption. To analyze these changes on living cells in real-time, a test system based on multiparametric chips with an array of sensors for monitoring pH and O(2) as well as electric impedance has been developed. Cells are cultivated on these chips and supplied with medium by a fluid perfusion set-up which mimics microphysiological conditions and allows for drug addition and removal. Human colon carcinoma cells LS174T were used as a model to test the effect of drugs. Cells growing on chips were monitored for 24 h and longer. Untreated cells showed a continuous increase in the rate of acidification, while the rate of respiration remained fairly constant. Addition of chloroacetaldehyde (50 microM) rapidly attenuated O(2) consumption with a gradual decrease in acidification following. In contrast, with cisplatin (16.7 microM) a delayed and gradual decrease in both the rates of acidification and respiration effect occurred over 2-3 days. These results provide insights to the mechanisms of action of these drugs, which are coherent with those already known. Thus, multiparametric sensor chips provide elementary information on drug action.
Asunto(s)
Antineoplásicos/farmacología , Técnicas Biosensibles , Proliferación Celular/efectos de los fármacos , Metabolismo Energético/efectos de los fármacos , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Línea Celular Tumoral , Ensayos de Selección de Medicamentos Antitumorales/instrumentación , Ensayos de Selección de Medicamentos Antitumorales/métodos , Electrónica Médica , Humanos , Consumo de OxígenoRESUMEN
An alternative assay for replacing animal experiments should serve the specific microphysiological needs of the cells and be endowed with multiparametric signal monitoring. These requirements are provided by a test system in which the key elements are biocompatible electronic sensor-chips. It is also connected to a medium perfusion set-up, which allows to control the supply of nutrients and test compounds, and the removal of culture medium. The chips are equipped with sensors that continuously monitor basic metabolic parameters and membrane-associated changes of living cells: pH-ISFETs for extracellular acidification, amperometric O2-sensors for oxygen consumption, and IDES for electrical impedance of the cell layer. Experiments with LS174T colon carcinoma cells in culture show the metabolic and electrical changes upon incubation with Zytochalasin B and chloroacetaldehyde. The signal patterns vary and indicate different mechanisms of action for these test compounds. With this test system it is possible to detect effects of unknown substances and mixtures, and to analyse the cellular probe for prolonged times.
Asunto(s)
Alternativas a las Pruebas en Animales , Técnicas Biosensibles/métodos , Células/química , Animales , División Celular , Línea Celular Tumoral , Células/citología , Células/metabolismo , Humanos , Ratones , Microcomputadores , Consumo de Oxígeno , Células Tumorales CultivadasRESUMEN
A constraint in the reliability of predictive chemosensitivity assays is linked to the fact that they analyze only a single cellular or biochemical parameter. A multiparametric test system using microsensor chips has been developed which can detect online microphysiological changes in living cells. Tumor cells were grown directly on glass- or silicon-based electronic sensor chips. Changes in extracellular pH and pO(2), reflecting metabolic activities, and changes in impedance, reflecting morphological properties, were monitored. In this study, colon and breast cancer cells as well as doxorubicin-sensitive and doxorubicin-resistant sarcoma cell lines were exposed to cytochalasin B, chloroacetaldehyde, or doxorubicin. Results show (1) reduction in medium acidification, (2) marked and rapid changes in O(2) consumption, and (3) modulations in impedance correlating with morphological changes observed in the microscope. Drug-resistant cells do not show these changes. Therefore, this microphysiological monitoring is a versatile tool for chemosensitivity testing of tumor cells.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias de la Mama/patología , Ensayos de Selección de Medicamentos Antitumorales/métodos , Modelos Teóricos , Sarcoma/patología , Neoplasias de los Tejidos Blandos/patología , Neoplasias de la Mama/metabolismo , Resistencia a Antineoplásicos , Humanos , Concentración de Iones de Hidrógeno , Nanotecnología/instrumentación , Consumo de Oxígeno , Reproducibilidad de los Resultados , Sarcoma/metabolismo , Neoplasias de los Tejidos Blandos/metabolismo , Células Tumorales CultivadasRESUMEN
Many different assays have been developed for testing the chemosensitivity of tumor cells in vitro, usually based on a single biochemical or cellular parameter. A multiparametric test system has been developed that accommodates on a single chip numerous sensors for metabolic parameters, deltapH and deltapO2, as well as for morphological changes. The cells grow directly on the chips and can be continuously monitored online up to several days. The effects of various chemotherapeutic drugs on the metabolic profile of several tumor cell lines have been investigated. In colon carcinoma-derived LS174T cells, cytochalasin B markedly increased oxygen consumption while decreasing the rate of extracellular acidification. These effects, which reflect the biochemical action of cytochalasin B, were reversible on drug removal. In contrast, chloroacetaldehyde markedly reduced respiration, which recovered when the drug was removed. Primary breast cancer cells also responded to chloroacetaldehyde with a marked reduction in deltapO2, followed by a reduced rate of acidification. Comparing the metabolism of doxorubicin-sensitive and -resistant mouse sarcoma S180 cells, the rates of acidification and respiration were inhibited by doxorubicin only in the sensitive cells, whereas in the resistant cells oxygen consumption even increased. These examples demonstrate that this chip-based test system provides rapid and important information for assessing chemosensitivity of tumor cells.