RESUMEN
The density-dependent growth inhibition of non-transformed cells may be associated with inefficient transduction of the proliferative signal from cell adhesion molecules. To verify this concept, the C3H10T1/2 fibroblasts were stably transfected with the gene coding for the fibronectin fragment III/10 (FNIII/10). This resulted in differences in gene's expression between original C3H10T1/2 cells and their FNIII/10 transfectants. No significant differences in growth properties were observed in the original or in the transfected cells. C3H10T1/2 cells and their transfectants, when co-cultured, displayed more cells at confluence than the cells cultured alone. Moreover, co-cultured C3H10T1/2 cells and their transfectants showed elevated levels of phospho-ERK1/2 compared to homogenous cultures. Results obtained indicate that cellular homogeneity is responsible for density-dependent growth inhibition.
Asunto(s)
Fibronectinas/genética , Transfección , Animales , Secuencia de Bases , Western Blotting , Técnicas de Cocultivo , ADN Complementario , Ratones , Ratones Endogámicos C3H , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Mensajero/genéticaRESUMEN
Two analogs of the peptide mimicking the 1977-1991 C- terminal part of fibronectin have been synthesized and tested. AWLI simulated human fibronectin fragment 1977-1991, whereas AWLII hybridized to both RGD and 1977-1991 fragments. AWLI and AWLII peptides inhibited the migration of the ovarian carcinoma cell line OVP10 regardless of the presence RGD. AWLI peptide inhibited spontaneous and fibronectin-activated cell migration and ERK1/2 activity. Neither AWLI nor fibronectin induced changes in FAK proteins, as could be judged from Western blots. In conclusion, it seems that the C-terminal fragment of fibronectin inhibits ERK1/2-dependent (random) migration of ovarian carcinoma cells.
Asunto(s)
Movimiento Celular/efectos de los fármacos , Fibronectinas/química , Proteína Quinasa 1 Activada por Mitógenos/antagonistas & inhibidores , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , Oligopéptidos/farmacología , Secuencia de Aminoácidos , Western Blotting , Adhesión Celular/efectos de los fármacos , Femenino , Fibronectinas/farmacología , Humanos , Proteína Quinasa 3 Activada por Mitógenos , Datos de Secuencia Molecular , Neoplasias Ováricas/patología , Fosforilación , Células Tumorales CultivadasAsunto(s)
Adenocarcinoma/metabolismo , Ascitis/metabolismo , Interleucina-8/metabolismo , Neovascularización Patológica , Neoplasias Ováricas/metabolismo , Adenocarcinoma/irrigación sanguínea , Adenocarcinoma/patología , Anciano , Animales , Ascitis/patología , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Persona de Mediana Edad , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Neoplasias Ováricas/irrigación sanguínea , Neoplasias Ováricas/patología , Células Tumorales CultivadasRESUMEN
In the present work we tested whether invasiveness of ovarian carcinoma cells could be considered as protein kinase C (PKC) dependent process. The migration and invasion studies were performed in Transwell chambers. Staurosporine, sphingosine and tamoxifen were used as PKC inhibitors. Also the effect of prolonged treatment with TPA was the subject of observation. The obtained results indicated that invasion understood as three step process (attachment, migration and matrix degradation) was affected by PKC inhibitors. The detailed studies, however, showed that attachment and matrix degradation ability of ovarian cancer cells was not changed by PKC inhibitors as opposed to migration which was, at least partly, regulated by protein kinase C.
Asunto(s)
Inhibidores Enzimáticos/farmacología , Neoplasias Ováricas/tratamiento farmacológico , Proteína Quinasa C/antagonistas & inhibidores , Anciano , Adhesión Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Colágeno , Regulación hacia Abajo , Femenino , Fibronectinas , Humanos , Invasividad Neoplásica , Metástasis de la Neoplasia , Neoplasias Ováricas/patología , Acetato de Tetradecanoilforbol/farmacología , Células Tumorales CultivadasRESUMEN
Murine sarcoma cell line (L-1) treated with promoting phorbol ester (TPA) showed decreased content and activity of protein kinase C (PKC) as measured by Western blotting and histone phosphorylation methods. The PKC depleted line (L-1R) produced bigger, tumors after s.c. transplantation into syngeneic mice and more spontaneous and artificial metastases developing after i.v. injection of tumor cells. The in vitro studies revealed decreased: adhesiveness, migratory and invasiveness properties of PKC depleted cells. Negative correlation between in vitro and in vivo studies were found.
Asunto(s)
Carcinógenos/farmacología , Sarcoma Experimental/tratamiento farmacológico , Sarcoma Experimental/patología , Acetato de Tetradecanoilforbol/farmacología , Animales , Western Blotting , Adhesión Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Femenino , Neoplasias Pulmonares/secundario , Ratones , Ratones Endogámicos BALB C , Trasplante de Neoplasias , Fosforilación , Proteína Quinasa C/efectos de los fármacos , Proteína Quinasa C/metabolismo , Sarcoma Experimental/enzimología , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
The invasion/migration of two cell lines, melanoma (MEW) and glioma (BT5C), and their counterparts treated for prolonged time with TPA were studied. On Western blots both cell lines expressed alpha, beta I protein kinase C isoforms, whereas beta II and gamma were not detected. The down-regulation of alpha and beta I PKC was observed in glioma cells after long treatment with TPA, whereas the same treatment of melanoma cells did not lead to PKC downregulation. The down-regulation of PKC was accompanied by stimulation of cell migration/invasion through Transwell chambers coated with collagen IV or fibronectin. In the case of melanoma cells treated with TPA, whose PKC was not down-regulated, the inhibition of migration/invasion was observed. The invasive properties of studied cells did not correlate with any conventional PKC isoform expression.
Asunto(s)
Carcinógenos/farmacología , Movimiento Celular/efectos de los fármacos , Isoenzimas/efectos de los fármacos , Invasividad Neoplásica , Proteína Quinasa C/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacología , Animales , Regulación hacia Abajo , Glioma/metabolismo , Glioma/patología , Humanos , Isoenzimas/metabolismo , Melanoma/metabolismo , Melanoma/patología , Proteína Quinasa C/metabolismo , Ratas , Células Tumorales CultivadasRESUMEN
Treatment of 10T1/2 cells with promoting phorbol ester drastically enhanced migration of the studied fibroblasts in a serum-supplemented medium. The same cells when exposed to ionomycin or TPA in a serum-free medium did not show any migration. The addition of 1% of serum induced spontaneous and TPA stimulated migration. Also EGF and PDGF separately or together induced the migration of 10T1/2 cells. Parallel studies of protein kinase C documented low enzymatic activity after treatment with TPA, whereas transcripts of PKC were shown independently of TPA treatment.
Asunto(s)
Carcinógenos/farmacología , Movimiento Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Proteína Quinasa C/biosíntesis , Acetato de Tetradecanoilforbol/farmacología , Animales , Movimiento Celular/fisiología , Células Cultivadas/efectos de los fármacos , Células Cultivadas/fisiología , Feto/efectos de los fármacos , Feto/enzimología , Fibroblastos/enzimología , Ratones , Ratones Endogámicos C3H , Reacción en Cadena de la Polimerasa , Proteína Quinasa C/análisisRESUMEN
Treatment with the tumor-promoting phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) and the second-stage promoter 12-O-retinoylphorbol 13-acetate induced down-regulation of protein kinase C (PKC) and enhanced the migration of C3H 10T1/2 cells. In the case of BT5C glioma cells the same negative correlation was observed only after treatment with TPA. The negative control 4 alpha-phorbol affected neither PKC activity nor the migratory ability of both cell lines.
Asunto(s)
Movimiento Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Glioma/fisiopatología , Ésteres del Forbol/farmacología , Animales , Línea Celular/efectos de los fármacos , Fibroblastos/enzimología , Glioma/enzimología , Glioma/patología , Ratones , Proteína Quinasa C/metabolismo , Ratas , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
The role of protein kinase C in migration of tumor cells from spheroid cultures was investigated using parental rat glioma cells and their TPA resistant counterparts. These two lines differed in their PKC content as judged by the histone phosphorylation method. Also 4 days of treatment with IRA led to PKC down-regulation. Cells having a drastically decreased PKC level migrated better than those having a normal PKC content.
Asunto(s)
Neoplasias Encefálicas/fisiopatología , Movimiento Celular , Glioma/fisiopatología , Proteína Quinasa C/metabolismo , Animales , División Celular/efectos de los fármacos , Línea Celular , Ratas , Acetato de Tetradecanoilforbol/farmacología , Células Tumorales CultivadasRESUMEN
Protein kinase C (PKC) from membrane and cytosol fractions of fast growing as well as confluent contact-inhibited cells was assayed by two methods: PDBu binding and histone phosphorylation. In cytosol and membrane fractions of fast growing cells and in the cytosol fraction of confluent cells a reasonable agreement between the two methods was found. In membranes of confluent cells good PDBu binding was not accompanied with histone phosphorylation. Also in contact-inhibited cells, serum-stimulated hydrolysis of phosphatidylinositol bisphosphate (PIP2) was found to be reduced.
Asunto(s)
Inhibición de Contacto , Fosfatos de Inositol/metabolismo , Ésteres del Forbol/metabolismo , Proteína Quinasa C/metabolismo , Animales , Células Cultivadas , Fibroblastos/enzimología , Ratones , Ratones Endogámicos C3H , Forbol 12,13-Dibutirato/metabolismo , Acetato de Tetradecanoilforbol/metabolismoRESUMEN
The effect of three retinoids, 13-cis-retinoic acid, arotinoid ethyl sulfone and retinal acetate, on methylcholanthrene (MCA) induced transformation of 10T1/2 cells was studied. It appears that 13-cis-retinoic acid and arotinoid ethyl sulfone prevent transformation in a direct way at the last stage of carcinogenesis. Retinal acetate, however, requires cell-to-cell contacts to inhibit the transformation of 10T1/2 cells.
Asunto(s)
Transformación Celular Neoplásica/efectos de los fármacos , Retinoides/farmacología , Animales , Células Cultivadas , Diterpenos , Metilcolantreno , Ratones , Ésteres de Retinilo , Tretinoina/farmacología , Vitamina A/análogos & derivados , Vitamina A/farmacologíaRESUMEN
The influence of 13-cis-retinoic acid on the light-caused injury of hematoporphyrin-pretreated nontransformed 10 T1/2 fibroblasts and N2a neuroblastoma cells was investigated. Retinoid reduced the percentage of damaged cells as assessed by the plating efficiency assay. N2a neuroblastoma cells were found to be better protected than 10 T1/2 cells by 13-cis-retinoic acid on hematoporphyrin light-induced injury.
Asunto(s)
Hematoporfirinas/farmacología , Fármacos Sensibilizantes a Radiaciones/farmacología , Tretinoina/farmacología , Animales , División Celular/efectos de los fármacos , División Celular/efectos de la radiación , Línea Celular , Células Cultivadas , Derivado de la Hematoporfirina , Isomerismo , Isotretinoína , Ratones , NeuroblastomaRESUMEN
Serum retinol levels were determined by a fluorometric method in patients with colorectal cancer or polyps and those with inflammatory bowel disease. Serum retinol levels in patients with benign or malignant colorectal polyps and stage B cancer (modified Dukes' classification) were similar to those found in controls. By contrast, serum retinol levels were significantly lower in patients with Dukes' stage C or D. Among cancer patients that were followed after surgical treatment serum retinol levels did not differ significantly from those found in controls. Patients who died of metastases during follow-up possessed very low serum retinol levels. These findings suggest that a decreased serum retinol level in cancer patients is a consequence rather than a precursor of the neoplastic process. Furthermore, this study suggests that the marked decrease in serum retinol level might be an indicator of poor prognosis in colorectal cancer patients after surgery.
Asunto(s)
Neoplasias del Colon/sangre , Pólipos del Colon/sangre , Neoplasias del Recto/sangre , Vitamina A/sangre , Adulto , Anciano , Anciano de 80 o más Años , Colitis Ulcerosa/sangre , Neoplasias del Colon/patología , Enfermedad de Crohn/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Neoplasias del Recto/patologíaRESUMEN
The aim of the study was to investigate the influence of the substances modifying cell phenotype (phorbol ester and 13-cis retinoic acid) on the growth of mouse neuroblastoma N2a cells in the mixed culture system. The results were compared with monoculture system and with tumor multicellular spheroids system where neoplastic cells are cultivated in the close contact from all sides. The suppression of the pleyotypic effect of phorbol ester was observed when N2a cells were cocultured with 3T3 fibroblasts. On the contrary, the close contact of neoplastic cells to each other (spheroids) do not change the stimulatory effect of phorbol ester on tumor cells proliferation. 13-cis retinoic acid suppressed neoplastic cells growth in all cell culture systems we used.
Asunto(s)
Acetato de Tetradecanoilforbol/farmacología , Tretinoina/farmacología , Células Tumorales Cultivadas/efectos de los fármacos , Animales , Comunicación Celular , Diferenciación Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Línea Celular , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Glioma/tratamiento farmacológico , Glioma/patología , Neuroblastoma/tratamiento farmacológico , Neuroblastoma/patología , Células Tumorales Cultivadas/patologíaRESUMEN
Protein kinase C (PKC) activity of cytosol and membrane fractions of 10T1/2 cells was studied. In cytosol of fast growing cells PKC activity was found in material eluted with 0.150 M NaCl whereas in membrane fractions activity was eluted with 0.065 and 0.150 M NaCl. In the membrane fraction of confluent cells, in contrast to cytosol, very low PKC activity was observed. The translocation pattern of PKC activity eluted with 0.065 M NaCl may be associated with proliferation.
Asunto(s)
Fibroblastos/citología , Proteína Quinasa C/metabolismo , Animales , División Celular , Línea Celular , Membrana Celular/enzimología , Citosol/enzimología , Fibroblastos/enzimología , Ratones , Ratones Endogámicos C3H , Concentración Osmolar , Cloruro de SodioRESUMEN
The aim of the study was to analyze the incidence of spontaneous thymic lymphomas in AKR mice kept on a diet with normal and excess retinoid content. The mice whose diet was supplemented with 13-cis-retinoic acid (250 mg per kg chow) developed less lymphomas than those kept on a standard diet (15 mg per kg chow). The effect of cyclophosphamide on the early stage of lymphomogenesis was tested using a single dose (100 mg per kg body weight), injected intraperitoneally to AKR mice. Increased incidence of lymphoma following cyclophosphamide administration was observed as result of a) low sensitivity of prelymphoma and lymphoma cells and/or b) immunosuppressive effect of cyclophosphamide.
Asunto(s)
Linfoma/inducido químicamente , Neoplasias del Timo/inducido químicamente , Tretinoina , Animales , Cocarcinogénesis , Ciclofosfamida/efectos adversos , Dieta , Femenino , Inyecciones Intraperitoneales , Isotretinoína , Linfoma/mortalidad , Masculino , Ratones , Neoplasias del Timo/mortalidadRESUMEN
In the mixed culture system nontransformed 3T3 fibroblasts cells inhibited the plating efficiency of neuroblastoma (N2a) cells, whereas heat or methanol killed fibroblasts did not. In contrast, chloroform/methanol extracted 3T3 cells inhibited the plating efficiency of neuroblastoma cells to the same extent as had been the case with living 3T3 cells. In the present study, we discuss the possibility that nontransformed cells surface components can modify the proliferation of neuroblastoma cells.
Asunto(s)
Comunicación Celular , Transformación Celular Neoplásica , Fibroblastos/citología , Neuroblastoma/patología , Absorción , Células Cultivadas , Cloroformo/farmacología , Cromatografía en Capa Delgada , Gangliósidos/análisis , Calor , Metanol/farmacologíaRESUMEN
Two pyrimidine analogs (metahalones) NY 3163, NY 3170 have been tested for their effects on the growth of rat glioma cells in monolayer and in spheroid culture. Both substances have earlier been found to inhibit the microtubule system in malignant cells. In glioma cells, arrest of mitosis was accompanied by repeated cycles of DNA synthesis, leading to different levels of polyploidization up to 16 and 32 ploid cells. The effect was not reversible through a culture period of 4 days. Reduced ability of directional migration of cells on a plastic surface was seen for 16 days after exposure. The reaction to microtubule inhibitors seems to differ depending on the type of tumour cells, where some malignant cells have earlier been reported to escape mitotic arrest and proceed to the next G1 phase, in contrast to the present glioma cells which undergo polyploidization.
Asunto(s)
Glioma/patología , Microtúbulos/efectos de los fármacos , Poliploidía , Pirimidinonas/farmacología , Animales , Recuento de Células , División Celular/efectos de los fármacos , Células Cultivadas , Replicación del ADN/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ratas , Factores de TiempoRESUMEN
Lung-conditioned medium (LCM) was obtained by incubation of BALB/c mouse lung tissue fragments in serum-free Eagle medium for 48 hours and subsequent separation by dialysis or chromatography on a Sephadex G-75 column. LCM fractions were tested for their ability to modulate proliferation of normal human endothelial cells and neoplastic cells (N2a, MCF, HEp-2) in vitro as assessed by plating efficiency and tritiated thymidine incorporation assays. It was found that LCM contained two kinds of factors, either stimulating (molecular weight: 50,000-70,000) or inhibiting (molecular weight: 12,000-20,000 and 3,000-5,000) cell proliferation.
Asunto(s)
División Celular , Sustancias de Crecimiento/fisiología , Pulmón/fisiología , Neoplasias/patología , Animales , Neoplasias de la Mama , Línea Celular , Cromatografía en Gel , Medios de Cultivo , Diálisis , Endotelio , Femenino , Inhibidores de Crecimiento/fisiología , Sustancias de Crecimiento/análisis , Humanos , Neoplasias Laríngeas , Pulmón/análisis , Ratones , Ratones Endogámicos BALB C , Peso Molecular , NeuroblastomaRESUMEN
Mice pretreated with cyclophosphamide (CY) had more tumor colonies in lungs following i.v. injection than their intact counterparts. CY treated mice restored with spleen cells showed incomplete reduction of the artificial metastasis in lungs. Conditioned media prepared from lungs contain two factors: stimulating and inhibiting cell proliferation in vitro. Conditioned media from CY treated mice were found to have a decreased inhibitory ability.