RESUMEN
In some neurological diseases, injury to neurones reflects an over-stimulation of their receptors for excitatory amino acids. This response may disturb the Ca(2+)-homeostasis and lead to a pronounced and sustained increase in the intracellular concentration of this ion. On the basis of data derived from correlative studies, calcium-binding proteins have been postulated to play a protective role in these pathologies. We tested, directly, the capacity of the three calcium-binding proteins calretinin (CR), calbindin D-28k (CB) and parvalbumin (PV) to buffer [Ca(2+)], and to protect cells against excitotoxic death. We used P19 murine embryonic carcinoma cells, which can be specifically induced (by retinoic acid) to transform into nerve-like ones. The differentiated cells express functional glutamate-receptors and are susceptible to excitotoxic shock. Undifferentiated P19-cells were stably transfected with the cDNA for CR, CB or PV, induced to differentiate, and then exposed to NMDA, a glutamate-receptor agonist. The survival rates of clones expressing CR, CB or PV were compared with those of untransfected P19-cells using the lactate-dehydrogenase assay. CR- and CB-expressing cells were protected from death during the first 2 h of exposure to NMDA. This protection was, however, transient, and did not suffice to rescue P19-cells after prolonged stimulation. Two of the three PV-transfected clones raised were vulnerable to NMDA-induced excitotoxicity; the third, which expressed the lowest level of PV, was protected to a similar degree as that found for the CR- and CB-transfected clones. Our results indicate that in the P19-cell model, CR and CB can help to delay the onset of cell death after excitotoxic stimulation.
Asunto(s)
Proteínas de Unión al Calcio/genética , Muerte Celular/fisiología , Enfermedades del Sistema Nervioso Central/tratamiento farmacológico , Sistema Nervioso Central/metabolismo , Neuronas/metabolismo , Fármacos Neuroprotectores/metabolismo , Neurotoxinas/metabolismo , Animales , Calbindina 2 , Calbindinas , Calcio/metabolismo , Proteínas de Unión al Calcio/metabolismo , Muerte Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Tamaño de la Célula/efectos de los fármacos , Tamaño de la Célula/fisiología , Sistema Nervioso Central/efectos de los fármacos , Sistema Nervioso Central/fisiopatología , Enfermedades del Sistema Nervioso Central/metabolismo , Enfermedades del Sistema Nervioso Central/fisiopatología , Agonistas de Aminoácidos Excitadores/farmacología , Aminoácidos Excitadores/metabolismo , Glicina/farmacología , Inmunohistoquímica , L-Lactato Deshidrogenasa/metabolismo , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Ratones , N-Metilaspartato/farmacología , Células Madre Neoplásicas/citología , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/metabolismo , Neuronas/efectos de los fármacos , Neuronas/patología , Parvalbúminas/genética , Parvalbúminas/metabolismo , Receptores de N-Metil-D-Aspartato/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/metabolismo , Proteína G de Unión al Calcio S100/genética , Proteína G de Unión al Calcio S100/metabolismo , Tretinoina/farmacologíaRESUMEN
Human mesothelioma cell lines were studied concerning the expression of the calcium-binding protein calretinin (CR), and the relation of the DNA index to their cell cycle. The results obtained for cell lines with different morphological characteristics, were compared to those from human mesothelial cells transfected with SV40 to escape senescence. Immunocytochemical expression of calretinin (confirmed by immunoblot) was observed in all mesothelioma cell lines but not in the control cells. It is suggested that calretinin is active in the first steps of carcinogenesis in all human mesotheliomas and during several stages in the evolution towards malignancy.
Asunto(s)
Ciclo Celular , Mesotelioma/metabolismo , Proteína G de Unión al Calcio S100/metabolismo , Western Blotting , Calbindina 2 , Línea Celular Transformada , Núcleo Celular/genética , Tamaño de la Célula , Transformación Celular Viral , ADN de Neoplasias/análisis , Citometría de Flujo , Humanos , Inmunohistoquímica , Mesotelioma/genética , Mesotelioma/patología , Índice Mitótico , Fenotipo , Proteína G de Unión al Calcio S100/inmunologíaRESUMEN
The expression of calretinin, a calcium-binding protein, has been studied in a series of 82 human colorectal adenocarcinomas. In 22.5% of the cases, part of the tumor cells were calretinin-positive, whereas the cells of the normal and paratumoral mucosa were always negative. Two types of cells from the tumoral mass reacted positively and selectively with calretinin-antisera: the tumor cells and giant fibroblasts. The neurons of enteric ganglia and reactive mesothelial cells also reacted positively to the same antibody. The results obtained by immunochemistry have been confirmed by Western blot analysis and in situ hybridization for calretinin mRNA. There is a correlation between the expression of calretinin and the degree of differentiation of the tumor. Well-differentiated tumors express calretinin in only 5% of the cases, whereas this percentage is 20% for moderately differentiated tumors and 66.6% for poorly differentiated or undifferentiated tumors. We conclude that calretinin is expressed by most undifferentiated colorectal adenocarcinomas, but only by a limited number of cells in well-differentiated tumors. The degree of its expression coincides also with additional signs of malignancy, such as an increase in the number of metastases in the regional lymph nodes and in other organs.
Asunto(s)
Adenocarcinoma/metabolismo , Colon/metabolismo , Neoplasias Colorrectales/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Proteína G de Unión al Calcio S100/metabolismo , Adenocarcinoma/secundario , Western Blotting , Calbindina 2 , Colon/patología , Neoplasias Colorrectales/patología , Células Epiteliales/metabolismo , Células Epiteliales/patología , Humanos , Técnicas para Inmunoenzimas , Hibridación in Situ , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Plexo Mientérico/citología , Plexo Mientérico/metabolismo , Proteínas del Tejido Nervioso/genética , ARN Mensajero/biosíntesis , Proteína G de Unión al Calcio S100/genética , Plexo Submucoso/citología , Plexo Submucoso/metabolismoRESUMEN
WiDr cells from a human colon adenocarcinoma cultivated in vitro express the calcium binding protein calretinin. The immunoreactivity is present in some interphasic cells and decreases after seven days in culture together with the augmentation of the cell number. Calretinin expression is maintained in the undifferentiated cells of the tumoral mass developed in nude mice and in recultivated isolated tumour cells from the xenograft. From the experiments here described, the protein expression is quantitatively influenced in vitro by the addition of drugs, such as colchicine and taxol, which intervene in cytoskeleton organisation. The percentage of the calretinin immunoreactive cells increases after the addition of colchicine to the medium while the immunoblot analysis shows a higher calretinin content in the cells treated with taxol.
Asunto(s)
Adenocarcinoma/metabolismo , Proteína G de Unión al Calcio S100/biosíntesis , Adenocarcinoma/patología , Animales , Western Blotting , Calbindina 2 , Adhesión Celular/efectos de los fármacos , Colchicina/farmacología , Citoplasma/metabolismo , Citoesqueleto/efectos de los fármacos , Humanos , Inmunohistoquímica , Ratones , Ratones Desnudos , Necrosis , Trasplante de Neoplasias , Paclitaxel/farmacología , ARN Mensajero/metabolismo , Factores de Tiempo , Células Tumorales CultivadasRESUMEN
Calretinin is a calcium-binding protein which is primarily expressed by certain cells of the nervous system, both central and peripheral. Its presence in nonexcitable cells has been little studied. Using a polyclonal antibody and paraffin sections we have analysed the presence of calretinin in the human ovary of different ages, and in polycystic ovaries. Our results revealed the selective presence of calretinin, specifically localised in the cells of the germinal epithelium, those of the theca interna and the theca lutein cells, in the cells of the neurovaso-epithelial association and of the canalicular structures of the mesovarium. Calretinin was also present in a few cells of the theca externa and some interstitial cells. No appreciable quantitative differences in the strength of the positive reaction were seen between the ovaries of different ages or between the normal and the polycystic ovaries. The presence of calretinin was confirmed by western blot analysis. The selective presence of calretinin in the human ovary, in androgenic cells and in the cells of the germinal epithelium is discussed in relation to its possible function as a Ca2+ buffer.
Asunto(s)
Proteínas Fetales/análisis , Ovario/química , Proteína G de Unión al Calcio S100/análisis , Adulto , Western Blotting , Calbindina 2 , Cuerpo Lúteo/química , Femenino , Humanos , Inmunohistoquímica , Ovario/embriología , Síndrome del Ovario Poliquístico/metabolismoRESUMEN
To elucidate the physiological role of the Ca2+ binding protein parvalbumin, we have generated transgenic mice carrying the full-length complementary DNA (cDNA) of rat parvalbumin under the control of the heavy-metal inducible metallothionein IIA promoter. Immunohistochemical and biochemical methods have been used to detect the presence of ectopic parvalbumin expression in different tissues. Here we show the expression of parvalbumin in endothelial cells lining the liver sinusoids in situ and after isolation in vitro. The hemodynamic effects of endothelin 1, a peptide hormone mediating potent vasoconstriction via calcium signalling, were investigated in the mouse liver perfused in situ. Vasoconstriction, thought to be mediated by the Ito cell, was not affected in the transgenic animals, whereas microvascular exchange, probed with the multiple indicator dilution technique, was markedly decreased in normal mice but virtually not affected in the transgenic animals. This suggests that ectopically expressed parvalbumin is involved in the regulation of Ca2+ signals in the sinusoidal endothelial cells. This animal model could be of interest to those working on the physiology of liver circulation.
Asunto(s)
Proteínas de Unión al Calcio/biosíntesis , Hígado/metabolismo , Parvalbúminas/biosíntesis , Animales , Proteínas de Unión al Calcio/genética , Endotelio/metabolismo , Endotelio/ultraestructura , Regulación de la Expresión Génica , Técnicas de Transferencia de Gen , Hígado/ultraestructura , Ratones , Ratones Transgénicos , Microscopía Inmunoelectrónica , Parvalbúminas/genética , RatasRESUMEN
We searched for the presence of calretinin (CR) in 12 colonic cancer cell lines using immunohistochemistry, Western blot analysis and the reverse transcriptase-polymerase chain reaction (RT-PCR). We were able to demonstrate that calretinin is expressed in the cell lines HT-29, WiDr, LoVo, LS180, CO112, CO115. SW480, SW620, COLO205 and SK-CO-1, while no detectable amounts were found in the cell lines SW1116 and Caco-2. In general, rapidly proliferating cell lines expressed calretinin, whereas the protein was absent from cell lines with a low multiplication rate. A possible role for calretinin in maintaining the proliferative cycle of tumor cells is discussed.
Asunto(s)
Adenocarcinoma/metabolismo , Neoplasias del Colon/metabolismo , Proteínas de Neoplasias/metabolismo , Proteína G de Unión al Calcio S100/metabolismo , Adenocarcinoma/patología , Western Blotting , Células CACO-2/metabolismo , Calbindina 2 , Neoplasias del Colon/patología , Humanos , Reacción en Cadena de la Polimerasa , Células Tumorales CultivadasRESUMEN
Calretinin is a Ca(2+)-binding protein of the EF-hand family which is expressed in colon adenocarcinomas and colon-derived tumor cell lines (e.g. WiDr), but is absent from normal human enterocytes. Its function has not as yet been elucidated, but some lines of evidence lead us to postulate its involvement in cell proliferation in these cells. In order to test whether calretinin is correlated with an undifferentiated, proliferating, or with a differentiated, state of cells, its expression was studied in the human colon adenocarcinoma clonal cell line HT29-18, which can be caused to differentiate into enterocyte-like cells by replacing glucose with galactose in the culture medium (glucose starvation differentiation). Treatment of HT29-18 cells with galactose led to a drop in the calretinin mRNA level and in protein expression as evidenced by immunocytochemical staining and Western blot analysis of cytosolic cell extracts. These results suggest that calretinin is present in HT29-18 cancer cells, mostly in those which are in the undifferentiated state. The possibility that calretinin is involved in maintaining the cells in an undifferentiated (cancerous) state is discussed.
Asunto(s)
Células HT29/citología , Proteína G de Unión al Calcio S100/análisis , Aminopeptidasas/análisis , Calbindina 2 , Muerte Celular , Diferenciación Celular , División Celular , ADN de Neoplasias/análisis , Galactosa , Glucosa , Células HT29/química , Células HT29/enzimología , Células HT29/ultraestructura , Humanos , Microvellosidades/ultraestructura , Índice Mitótico , Proteínas de Neoplasias/análisis , ARN Mensajero/análisis , Proteína G de Unión al Calcio S100/fisiologíaRESUMEN
The colon adenocarcinoma cell line WiDr expresses the calcium-binding protein calretinin (CR). In order to deduce possible functions of calretinin in these cells we decreased its concentration by antisense techniques. Treatment of WiDr cells with phosphorothioate antisense oligodeoxynucleotides (AS-ODNs) led to a drop in calretinin expression, as evidenced by immunohistochemical staining of WiDr cells and Western blot analysis of cytosolic cell extracts. The morphology of these epithelial cells changed from polygonal to spherical and they formed dense cell clusters. Cells displaying morphological alterations typical for apoptotic cells were observed after incubation with AS-ODNs, as evidenced by phase-contrast and electron microscopy. The mitotic rate of AS-ODN-treated cells dropped significantly, as demonstrated by mitotic labeling and time-lapse microcinematography. Furthermore, an accumulation of cells in phase G1 and a reduction of [3H]thymidine-labeled cells was observed in antisense-treated cells. The basal level of [Ca2+]i was not influenced by the down-regulation of calretinin. WiDr cells incubated with the nonsense, reverse-sense, or with an oligodeoxynucleotide with a totally unrelated sequence did not show any significant differences when compared to control cells. We conclude that calretinin levels have an impact on the progression of the cell cycle of WiDr cells.
Asunto(s)
Células CACO-2/citología , Oligonucleótidos Antisentido/farmacología , Proteína G de Unión al Calcio S100/metabolismo , Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Western Blotting , Células CACO-2/fisiología , Calbindina 2 , Calcio/metabolismo , Ciclo Celular/fisiología , División Celular/efectos de los fármacos , División Celular/fisiología , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo/fisiología , Humanos , Inmunohistoquímica , Cinética , Proteínas del Tejido Nervioso/metabolismo , Proteína G de Unión al Calcio S100/genéticaRESUMEN
In a series of 23 cases of mesothelioma of either the epithelial, sarcomatoid or the mixed type, the expression of three calcium-binding proteins (calretinin, parvalbumin and calbindin-D28k) was studied using immunohistochemical techniques on paraffin sections. The results show that calretinin is expressed in mesotheliomas of the epithelial type (papillary, adenomatous or solid) and by the epithelial component of the mixed tumours. The immunohistochemical reaction is specific and reproducible. The tissues of the pulmonary parenchyma and of the pleura are negative for calretinin except for the rare fibroblasts and some skeletal muscle fibres situated in the interstices of, or near the epithelial tumour mass. The sarcomatoid mesotheliomas and the sarcomatoid component of the mixed tumours do not express calretinin. Parvalbumin and calbindin-D28k are expressed neither in mesotheliomas nor in normal lung tissue. Primary adenocarcinomas of the lung are negative for all three calcium binding proteins cited. Thus, calretinin seems to represent a selective marker for mesotheliomas of the epithelial type and allows their differentiation from metastases of lung adenocarcinomas.
Asunto(s)
Biomarcadores de Tumor/análisis , Mesotelioma/metabolismo , Neoplasias Pleurales/metabolismo , Proteína G de Unión al Calcio S100/análisis , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Anciano , Anciano de 80 o más Años , Calbindina 1 , Calbindina 2 , Calbindinas , Diagnóstico Diferencial , Epitelio/patología , Femenino , Humanos , Inmunohistoquímica , Neoplasias Pulmonares/patología , Masculino , Mesotelioma/patología , Persona de Mediana Edad , Parvalbúminas/análisis , Neoplasias Pleurales/patología , Neoplasias Pleurales/secundarioRESUMEN
Parvalbumin is thought to act as a Ca2+ buffer in skeletal muscle fibers, but its physiological role in brain, kidney, and testis remains unclear. We have transfected parvalbumin cDNA into a human ovarian adenocarcinoma cell line, which normally does not express this protein. The induced expression of parvalbumin under the control of three different promoters causes: (1) changes in the morphology of the cells from epitheloid to fusiform, (2) an increase in motility of whole cell clusters, and (3) a decrease in the mitotic rate. Transfection with a mutated cDNA of rat parvalbumin incapable of binding Ca2+ had no effect on these three parameters. Our results indicate that ectopic expression of parvalbumin influences not only cell division [Rasmussen and Means (1989) Mol. Endocrinol. 3, 588-596], but also cell shape and motility by modulating intracellular Ca2+ handling. This may be a basic function of parvalbumin when it is intrinsically expressed in differentiated nonmuscle cells.
Asunto(s)
Técnicas de Transferencia de Gen , Ovario/citología , Parvalbúminas/genética , Línea Celular Transformada , Movimiento Celular , Tamaño de la Célula , ADN Complementario/genética , Femenino , Humanos , Ovario/metabolismo , Parvalbúminas/metabolismoRESUMEN
Transgenic mice expressing rat parvalbumin under the control of the human metallothionein IIA (MTII A), SV-40 early, and neuron-specific enolase (NSE) promoters were produced. Ectopic expression was analyzed by RNA polymerase chain reaction and RNase protection in combination with immunohistochemistry. From a total of 25 transgenic lines 18 were found to express the transgene. Expression strength and tissue specificity were dependent upon the promoter used and varied considerably among animal lines produced with the same construct. Highest constitutive MT IIA-driven expression was found in lung, liver, heart, and kidney, as well as in brain, and lower amounts of transgene expression were found in spleen, testis, and muscle. Immunohistochemistry of tissue sections of metallothionein-parvalbumin transgenic strain 29 in the non-induced state revealed that ectopic PV mRNA is translated into protein. Short-term induction of the MT IIA promoter by CdSO4 or CdCl2 leads to a shift in tissue specificity and does not increase ectopic expression in tissues where the transgene is active in the noninduced state. As expected the NSE promoter showed highest activity in brain. However, NSE-driven expression could also be detected to various degrees in all investigated tissues. SV-40-dependent PV expression showed no tissue preference and varied considerably among different strains. Except for the observation that the SV-40-PV construct showed lower yields in transgenic production and reduced numbers of positive offspring no obvious impairment of growth or behavior as a consequence of transgenic PV expression could be detected.
Asunto(s)
Ratones Transgénicos/metabolismo , Parvalbúminas/biosíntesis , Animales , Secuencia de Bases , Línea Celular Transformada , ADN Complementario , Técnicas de Transferencia de Gen , Humanos , Inmunohistoquímica , Ratones , Datos de Secuencia Molecular , Especificidad de Órganos , Parvalbúminas/genética , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas , ARN/análisisRESUMEN
Calretinin, a cytosolic calcium-binding protein, is widely expressed in mammalian and chicken neurons, including subpopulations of dorsal root ganglion neurons. In chicken embryo spinal ganglion cells, calretinin first appears on the 9th day of incubation. To determine whether the expression of this protein is maintained in vitro and depends on the developmental stage, dissociated dorsal root ganglion cultures from chick embryos at E6 and E10 (before or after establishing connections) were processed for calretinin immunohistochemistry. Cultured neurons from E6 embryos never showed calretinin immunoreactivity at any culture duration, whereas neurons from E10 embryos displayed strong immunostaining immediately after attaching to the culture dish. Quantitative evaluation revealed that the percentage of calretinin-positive neurons increased until day 3 in culture and afterwards declined in parallel the total cell number. These results indicate that primary sensory neurons express calretinin in vitro similarly as in vivo and the gene expression depends from the establishment of connections with peripheral targets.
Asunto(s)
Ganglios Espinales/química , Neuronas/química , Proteína G de Unión al Calcio S100/análisis , Animales , Biomarcadores/química , Calbindina 2 , Células Cultivadas , Embrión de Pollo , Ganglios Espinales/citología , InmunohistoquímicaRESUMEN
We used an atomic force microscope (AFM) to image samples immersed in a fluid in order to study the dynamic behavior of the membranes of living cells. AFM images of cultured cells immersed in a buffer were obtained without any preliminary preparation. We observed surface changes and displacements which suggest that the cells were still alive during the measurements. Some membrane details imaged with the AFM have also been observed using a scanning electron microscope and their dynamic behavior has been confirmed by microcinematography. We believe that the AFM will offer new insights into the exploration of dynamic changes affecting cell membranes.
Asunto(s)
Células/ultraestructura , Microscopía/métodos , Fenómenos Biofísicos , Biofisica , Línea Celular , Técnicas Citológicas , Estudios de Evaluación como Asunto , Humanos , Microscopía/instrumentación , Microscopía Electrónica de RastreoRESUMEN
Ca2+ ions intervene during different phases of the progression of the cell cycle, but only one calcium-binding protein, calmodulin, has been shown to be associated with dividing cells. We therefore screened cancer cells for the presence of other related calcium-binding proteins. Using molecular biological and immunohistochemical techniques we show that human tumor cells of epithelial origin, express calretinin. Calretinin immunoreactivity can be demonstrated at precise moments of the cell cycle and, in particular, in phase G1 and during mitosis. During mitosis calretinin is localized both in the cytoplasm and in the mitotic spindle. In the cytoplasm we find calretinin after prophase and until telophase. In the spindle apparatus, calretinin is already present in cells in prometaphase and persists in all the succeeding mitotic phases. It is associated with the kinetochore microtubules but, in contrast to calmodulin, also with the polar microtubules. The role that calretinin plays in well-defined moments of the cell cycle of these cells is as yet unknown, but our results strongly suggest that, in collaboration with other molecules, calretinin intervenes in the dynamic phenomena regulating the separation of the chromosomes.
Asunto(s)
Calcio/metabolismo , Ciclo Celular/fisiología , Proteína G de Unión al Calcio S100/biosíntesis , Adenocarcinoma , Calbindina 2 , Clonación Molecular , Humanos , Inmunohistoquímica , Cinética , Proteína G de Unión al Calcio S100/genética , Transducción de Señal , Células Tumorales CultivadasRESUMEN
Clonogenic cultures from 27 lung cancers were realised. For 68% of cases a clonogenic growth was observed, but no direct correlation could be done with the differentiation of the tumor or with the presence of ganglion metastases. EGF in medium increased the number of colonies obtained in 60% of cases.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/patología , Carcinoma de Células Pequeñas/patología , División Celular/fisiología , Neoplasias Pulmonares/patología , Células Tumorales Cultivadas/patología , Ensayo de Tumor de Célula Madre , Biomarcadores de Tumor/análisis , Factor de Crecimiento Epidérmico/análisis , Humanos , Pulmón/patología , PronósticoRESUMEN
"In vivo" and "in vitro" morphological analysis of associations of cells ("rosettes") involved in immune response in human tumoral effusions revealed the existence of cell interactions either by simple membrane apposition between the cell projections or by gap-like junctions between two adjacent cells; endocytotic phenomena were also observed. The giant fibroblastic cells seen "in vitro" ("myofibronoblasts") reacting positively to anti-human macrophage Mabs, might be the cells presenting antigen to lymphocytes.
Asunto(s)
Líquido Ascítico/patología , Carcinoma/patología , Linfocitos/patología , Macrófagos/patología , Neoplasias Ováricas/patología , Anciano , Anciano de 80 o más Años , Comunicación Celular , Endocitosis , Femenino , Humanos , Uniones Intercelulares/ultraestructura , Persona de Mediana Edad , Células Tumorales Cultivadas/patologíaRESUMEN
The cells from a malignant fibrous histiocytoma were enzymatically isolated and cultured in vitro. The cultures were observed by microcinematography and with an electron microscope after fixation and embedding. The interactions between histiocytic and tumor cells resulted in tumor cell death. The microcinematographic study revealed that filiform projections of the histiocytic cell protrude toward the tumor cell surface making contacts for varying periods of time. The tumor cell then contracts and almost simultaneously some granules (lysosomes?) appear in the tumor cell cytoplasm. These granules eventually fuse with each other thus increasing in volume. The resulting granulation heads for the periphery of the tumor cell at a site previously touch by a projection of a histiocytic cell; at this site the membrane bursts and the tumor cell dies leaking amorphous cytoplasmic material. Before bursting the tumor cell does not show any noticeable morphological changes.
Asunto(s)
Histiocitoma Fibroso Benigno/patología , Supervivencia Celular , Gránulos Citoplasmáticos/ultraestructura , Femenino , Histiocitoma Fibroso Benigno/ultraestructura , Humanos , Microscopía Electrónica , Persona de Mediana Edad , Películas Cinematográficas , Células Tumorales Cultivadas/citología , Células Tumorales Cultivadas/ultraestructuraRESUMEN
We expose here our first results concerning clonogenic growth of 40 cases of malignant tumor plated in semi-solid medium (methylcellulose). The growth is determined by the presence at day 21 of colonies (cellular group with more than 60 micron diameter). 26 cases were considered significant. A 61% growth has been observed with primary solid tumors, and 83.4% with ascites, ovarian carcinoma growing the best. Cellular viability, plating efficiency and when the cellular number allowed it, linear graphic curves have been established. An ultrastructural morphological study, illustrated here by 2 cases--one ovarian carcinoma and one colonic adenocarcinoma cell line--has shown that all colony 's cells belong to a same cellular type and present various degrees of differentiation, suggesting a possible analogy between our observations and the hypothetical tumor cell model, concerning human carcinoma's organisation and self renewal.