RESUMEN
The development of transformed cell lines evolving from an embryo fibroblastic C3H primary culture was followed before and after the ageing crisis using different techniques. By flow cytometry, alteration of subpopulations having different DNA content and altered metabolic activity was observed after the crisis, with the trend to assume a near tetraploid DNA index at higher passages. The fibrin clot retractile activity was lost in all cases during the ageing crisis, but the outcome did not present uniform values of growth characteristics or chromosome number and tumorigenicity appeared to be a nonstable property of the transformed cell lines.
Asunto(s)
Línea Celular Transformada/citología , Animales , División Celular , Línea Celular Transformada/metabolismo , Transformación Celular Neoplásica/patología , ADN/metabolismo , Embrión de Mamíferos/citología , Fibrina/metabolismo , Fibroblastos/citología , Citometría de Flujo , Cariotipificación , Ratones , Ratones Endogámicos C3H , ARN Bicatenario/metabolismoRESUMEN
The growth inhibitory effects, the reduction of [3H]-TdR incorporation and the perturbation of the cell cycle induced by the new agent mitozolomide on the M14 human melanoma cell line and on the SW626 human ovarian cancer cell line were compared to those produced by BCNU. Flow cytometry showed an interesting difference: at the high concentration mitozolomide induced an accumulation of cells in S middle and S late-G2-M phase of the cell cycle whereas BCNU caused only a block in S late-G2-M. Further studies were aimed at investigating the susceptibility of freshly isolated human ovarian cancer cells to pharmacologically reasonable mitozolomide concentrations. Only in one out of 16 primary cultures of human ovarian cancers was mitozolomide able to induce cell cycle perturbation, suggesting that ovarian carcinoma cells may not be sensitive to this drug.
Asunto(s)
Antineoplásicos/farmacología , Compuestos de Mostaza Nitrogenada/farmacología , Anciano , Carmustina/toxicidad , Ciclo Celular/efectos de los fármacos , Línea Celular , Relación Dosis-Respuesta a Droga , Femenino , Citometría de Flujo , Humanos , Melanoma/metabolismo , Persona de Mediana Edad , Mitosis/efectos de los fármacos , Neoplasias Ováricas/metabolismo , Timidina/metabolismoRESUMEN
Mitozolomide is one of the most effective drugs against Lewis lung carcinoma in the mouse. Two IP doses of 40 mg/kg (days 6 and 15 after IM transplantation of 3LL) or four doses of 20 mg/kg given at various intervals (starting from day 6) increased survival time by 100%. A single IP dose of 80 mg/kg was toxic, and 10 mg/kg was ineffective even when this dose was given on eight occasions. The pharmacokinetics of mitozolomide was investigated in 3LL-bearing mice by HPLC assay. Peak drug levels were achieved in tumor 15 min after IP treatment, after which they declined according to first-order kinetics, with a half-life of 80-100 min (the same as in plasma). No dose-dependent kinetics was observed. Flow cytometry studies showed an accumulation of 3LL cells in G2M 24 h after drug treatment. This cell cycle perturbation was reversed 96 h after the inactive dose of 10 mg/kg, but not after the effective dose of 40 mg/kg.
Asunto(s)
Neoplasias Pulmonares/metabolismo , Compuestos de Mostaza Nitrogenada/metabolismo , Animales , Ciclo Celular , Evaluación Preclínica de Medicamentos , Citometría de Flujo , Semivida , Cinética , Neoplasias Pulmonares/tratamiento farmacológico , Masculino , Ratones , Ratones Endogámicos C57BL , Compuestos de Mostaza Nitrogenada/uso terapéutico , Análisis de RegresiónRESUMEN
Out of 130 ovarian cancer patients the DNA index of cells from ovarian carcinoma was studied in 56 cases in which cytospin preparations showed the presence of atypical cells. In 24 patients the population had a diploid DNA index (1.0) and in the others the DNA index ranged from 1.2 to 2.0 (tetraploid). No hypodiploid or hypertetraploid populations were detected. Repeated samples from the same patients did not show any significant differences and primary culture did not alter the DNA index. In contrast, cell cycle phase distribution differed greatly from sample to sample, as also the ratio between DNA diploid and DNA aneuploid populations. Primary culture was successful in 57% of the tumours, with a higher percentage of success in DNA aneuploid tumours. After primary culture the ratio between DNA aneuploid cells and DNA diploid cells increased. In relation to the histological gradings of malignancy, DNA aneuploid cells clustered in the highest grade of malignancy. The mean S-phase for tumours with a DNA index of 1.0 was 3.5 and 14.1% for those with DNA index greater than 1. Ovarian carcinomas show a large difference in DNA index between patients even after primary culture.
Asunto(s)
ADN de Neoplasias/análisis , Neoplasias Ováricas/análisis , Femenino , Citometría de Flujo , Humanos , Técnicas In Vitro , Interfase , Mitosis , Neoplasias Ováricas/patología , Ploidias , Factores de TiempoRESUMEN
The presence of fibrin deposits in the microenvironment of tumor cells has been reported repeatedly and considered to play an important role in tumor biology. Among the mechanisms by which fibrin may be deposited in tumors, procoagulant activities (PCA) of different types have been described in cancer cells. The present study was aimed at establishing whether the nature of cellular PCA was a characteristic associated with malignant transformation. PCA of normal and transformed cells was investigated on pairs of murine and human origin. The transformed counterparts were obtained after treatment with low-dose radiation, chemical carcinogen, viral infection or after in vitro spontaneous immortalization. Both before and after any type of transformation cell PCA was of the tissue thromboplastin type, identified on the basis of biological criteria: requirement of factor VII for its expression and lack of inhibition by the serine protease inhibitor diisopropylfluorophosphate (DFP). Transformed cells of murine origin showed significantly lower activity than their normal counterparts, whereas all the transformed human cell lines expressed significantly higher activity than normal. An inverse correlation between the levels of PCA and the cell density in culture was observed in all but one of the lines tested. These findings suggest that the factor X activating property described in some tumors or in transformed cells cannot be considered as a general marker of transformation.
Asunto(s)
Coagulación Sanguínea , Transformación Celular Neoplásica , Transformación Celular Viral , Animales , Coagulación Sanguínea/efectos de los fármacos , Coagulación Sanguínea/efectos de la radiación , Pruebas de Coagulación Sanguínea , Recuento de Células , Línea Celular , Transformación Celular Neoplásica/efectos de los fármacos , Transformación Celular Neoplásica/efectos de la radiación , Transformación Celular Viral/efectos de los fármacos , Transformación Celular Viral/efectos de la radiación , Células Cultivadas , Humanos , Ratones , Ratones Endogámicos C3HRESUMEN
Mouse fibroblasts were cultured by three different procedures: (a) changing the 0.2 ml/cm2 of growth medium every 2nd d and seeding 1 X 10(5) cells/cm2 after confluency; (b) changing the 0.4 ml/cm2 of growth medium only at subculture performed at confluency by a 1:2 split and keeping the bottles incubated on a rocking platform; (c) the same as Method b but keeping the bottles stationary throughout culture. By Method a no lines were immortalized over 36 experiments whereas Method b gave 1/4 immortalized lines and Method c gave 10:12 immortalized lines. Cells always went into crisis at the 9th to 11th doubling. Immortalized lines had a tetraploid DNA content.
Asunto(s)
Línea Celular , Ratones Endogámicos C3H , Animales , Ciclo Celular , Retracción del Coagulo , Medios de Cultivo , Fibroblastos/citología , RatonesRESUMEN
Resistance of mouse M5076 (M5) ovarian reticular cell sarcoma to cyclophosphamide (CTX) was obtained in vivo by repeated drug treatment followed by transplantation of the regrowing tumor. After 16 passages, we obtained an M5 subline resistant to CTX (M5-CTX-16R). Median survival times were approximately 29 and 39 days for M5 and M5-CTX-16R, respectively. Survival of M5-bearing mice given a single i.p. dose of 200 or 300 mg/kg was 160 and 168% of controls, respectively, whereas in M5-CTX-16R it ws 103 and 123%, respectively. The resistance was not reversible after 14 additional passages with no further CTX treatment. M5 and M5-CTX-16R appear similar in histological features, pattern of metastasis formation, and DNA content, as assessed by flow cytometry (hypotetraploid). Metastases of M5-CTX-16R were also resistant to CTX. Flow cytometry studies 12 and 24 hr after CTX treatment revealed a block in S and G2-M phases in both tumors. After 48 hr and at subsequent times, no cytokinetic pertubation was evident in M5-CTX-16R, whereas in M5 marked accumulation of cells in G2-M was observed at 48, 72, 96, and 120 hr. Cross-resistance was found between CTX, L-phenylalanine mustard, chlorambucil, and hexamethylmelamine. M5-CTX-16R was sensitive, but less so than M5, to cis-platinum, 1,3-bis(2-chloroethyl)-1-nitrosourea, and imidazole-4-carboxamide,5-(3,3-dimethyl-1-triazene). Adriamycin was equally active on M5 and M5-CTX-16R, while 4'-demethylepipodophyllotoxin-9-(4,6-O-ethylidine-beta-D-glucopyranoside) was inactive. This model appears to be suitable for studies on the mechanism of resistance to CTX and alkylating agents and for screening new, non-cross-resistant drugs.
Asunto(s)
Antineoplásicos/uso terapéutico , Ciclofosfamida/uso terapéutico , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Neoplasias Ováricas/tratamiento farmacológico , Animales , Carmustina/uso terapéutico , Ciclo Celular/efectos de los fármacos , Línea Celular , Dacarbazina/uso terapéutico , Resistencia a Medicamentos , Femenino , Linfoma de Células B Grandes Difuso/fisiopatología , Melfalán/uso terapéutico , Ratones , Ratones Endogámicos C57BL , Neoplasias Ováricas/fisiopatologíaRESUMEN
Normal and established human epithelial cell lines obtained from the same organs were compared for their capacity to retract a fibrin clot. Fibrin clot retraction was maximal in normal epithelial cells, reduced in established nontumorigenic lines, and lost in tumorigenic cancer cell lines. Fibrin clot retraction efficiency seemed to be related to the degree of cellular spreading within the clot at the end of the test. Previous works and the present study suggest that fibrin clot retraction is correlated with some steps of cell transformation in vitro.
Asunto(s)
Transformación Celular Neoplásica , Fibrina , Neoplasias/patología , Comunicación Celular , Recuento de Células , Línea Celular , Retracción del Coagulo , Células Epiteliales , Fibrina/farmacología , HumanosRESUMEN
Of 89 samples of cancer cells from ovarian cancer patients primary cultures representative of the cancer cell population could be established in 17. The clinical response to polychemotherapy was studied in relation to the inhibition of thymidine uptake by the cultured cells. Cultures of each patient's tumour were exposed to concentrations of the drugs the patients had been given for long enough to reproduce the area under the curve (AUC) of the plasma levels resulting from in vivo dosage. Full agreement was observed between the degree of thymidine uptake inhibition induced by at least one of the drugs administered to the cultured cells and the degree of clinical response. This approach may prove useful in pharmacological studies as a means of obtaining ovarian cancer cell populations representative of human tumours, even though the number of tumours that can be successfully evaluated in vitro is still too small to serve as a sound basis for prediction.
Asunto(s)
Antineoplásicos/uso terapéutico , Neoplasias Ováricas/tratamiento farmacológico , Adulto , Anciano , Altretamina/farmacología , Altretamina/uso terapéutico , Antineoplásicos/farmacología , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Cisplatino/farmacología , Cisplatino/uso terapéutico , Ciclofosfamida/farmacología , Ciclofosfamida/uso terapéutico , Relación Dosis-Respuesta a Droga , Doxorrubicina/farmacología , Doxorrubicina/uso terapéutico , Quimioterapia Combinada , Femenino , Fluorouracilo/farmacología , Fluorouracilo/uso terapéutico , Humanos , Persona de Mediana Edad , Neoplasias Ováricas/patologíaRESUMEN
Fibroblast-like cells derived from C3H mice at the first passage in culture, and a line from the same origin, which had undergone spontaneous transformation at the eleventh passage, were seeded on fibrin plates and tested for their attachment and spreading. After 4-24 hours normal cells had a spider-like morphology, while transformed cells remained in round clusters, flattened on the substrate. This data represent the morphological counterpart of the abnormal fibroblast-fibrin interaction shown in a tridimensional model, where transformed cells were found unable to induce the retraction of a fibrin clot (FCR).
Asunto(s)
Transformación Celular Neoplásica , Fibroblastos/citología , Animales , Adhesión Celular , Movimiento Celular , Células Cultivadas , Fibrina , RatonesRESUMEN
The expression of transformation parameters (inhibition of cell division during cell crowding, anchorage dependence, loss of fibrin clot retractile activity and secretion of plasminogen activator) was studied in a heterospecific cellular hybrid, made between established L(TK-) cells and the normal human MRC-5 cells. The hybrid nature of the cross was confirmed by the ability to incorporate [3H]-thymidine, by growth in selective HAT medium, by the identification of human chromosomes and by the expression on the surface of 100% of hybrid cells of a human glycoprotein, which is recognized by the 4F2 monoclonal antibody. The hybrid cultures showed cell cycle inhibition which became less stringent with increasing population doublings and the loss of human chromosomes. Fibrin clot retraction and anchorage dependence were absent in spite of the presence of many human chromosomes. The two properties were present or lost simultaneously in the normal parent cells and in the transformed parent or hybrid cells respectively. The human type of plasminogen activator was secreted even with very little human genetic material left, and a complete dissociation between fibrin clot retraction and production of plasminogen activator was observed. The data strengthen the hypothesis that transformation is a multistep process that involves complex genetic control and where cells progressively express different phenotypes and escape growth control.
Asunto(s)
Coagulación Sanguínea , Células Híbridas/inmunología , Activadores Plasminogénicos/análisis , Animales , División Celular , Fusión Celular , Línea Celular , Transformación Celular Neoplásica , Células Cultivadas , Fibrina , Glicoproteínas/inmunología , Humanos , Células Híbridas/citología , Cariotipificación , Cinética , RatonesRESUMEN
Two sublines of Walker 256 carcinoma have been characterized for their ability to metastasize and to induce cachexia. The invasive, metastasizing line A induced terminal anorexia in rats with a mean survival time of 27 +/- 1.5 days. The non-invasive line B induced early anorexia and cachexia with a mean survival time of only 15 +/- 1 days. At death, the line B tumor was still smaller than the line A one, and no metastases were detectable. These two sublines are discussed as a composite model for studying anorexia and cachexia together with invasion and metastasis.
Asunto(s)
Carcinoma 256 de Walker/patología , Animales , Anorexia/etiología , Peso Corporal , Caquexia/etiología , Carcinoma 256 de Walker/complicaciones , Carcinoma 256 de Walker/fisiopatología , Línea Celular , Ingestión de Líquidos , Ingestión de Alimentos , Riñón/fisiopatología , Metástasis de la Neoplasia , RatasRESUMEN
The two dosage schedules of VP16 that gave the least and the greatest efficacy in Lewis lung carcinoma of the mouse were selected for evaluation of the cytokinetic effects observable in vivo at different intervals after treatment (schedule A: 40 mg/kg IV, on day 8 after transplant; schedule B: 13 mg/kg IV, repeated on days 8, 11, and 14 after transplant). After the single dose and after each repeated dose there was a marked increase in the percentage of cells in the LS-G2-M phases, with a corresponding decrease in the percentage of cells in G0-G1. The number of neoplastic tetraploid cells compared with normal diploid cells in the tumor was reduced after the single IV dose, and more markedly so after repeated doses. This study suggests that the more marked delay of cancer cell growth and greater effectiveness observed with schedule B is related to repeated blockage of the LS-G2-M phases.
Asunto(s)
ADN de Neoplasias/análisis , Etopósido/uso terapéutico , Citometría de Flujo , Neoplasias Pulmonares/tratamiento farmacológico , Podofilotoxina/análogos & derivados , Animales , Ciclo Celular/efectos de los fármacos , Línea Celular , Neoplasias Pulmonares/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Trasplante de Neoplasias , Neoplasias Experimentales/tratamiento farmacológico , Neoplasias Experimentales/metabolismoRESUMEN
In a double-blind trial the effect of ICRF 159 or placebo associated with radiotherapy was compared in 70 patients with cervical carcinoma. No significant differences were found on histopathological examination of surgical specimens obtained 6 weeks after the end of the treatment. Clinical evaluation after a follow-up of 46 +/- 5 months, also showed no significant effect of ICRF 159 with radiotherapy.
Asunto(s)
Carcinoma de Células Escamosas/radioterapia , Piperazinas/uso terapéutico , Razoxano/uso terapéutico , Neoplasias del Cuello Uterino/radioterapia , Carcinoma de Células Escamosas/tratamiento farmacológico , Ensayos Clínicos como Asunto , Método Doble Ciego , Femenino , Humanos , Estadificación de Neoplasias , Placebos , Razoxano/efectos adversos , Neoplasias del Cuello Uterino/tratamiento farmacológicoAsunto(s)
Neuronas/metabolismo , Serotonina/metabolismo , Animales , Línea Celular , Clomipramina/farmacología , Femenino , Fluoxetina/farmacología , Cinética , Ratones , EmbarazoAsunto(s)
Retracción del Coagulo , Fibroblastos/fisiología , Neoplasias/fisiopatología , Adolescente , Adulto , Línea Celular , Supervivencia Celular , Niño , Embrión de Mamíferos , Femenino , Fibrina/fisiología , Humanos , Recién Nacido , Masculino , Embarazo , Piel/fisiopatología , Fenómenos Fisiológicos de la PielRESUMEN
VP 16 activity was investigated on cells derived from primary tumor or metastases of Lewis lung carcinoma grown in vitro. After 24 hr exposure to VP 16 we found a marginal effect at 0.1 microgram/ml, but at 1 and 10 microgram/ml [3H]-TdR uptake was 37 and 85% less than controls for primary tumor and 43 and 83% for metastases; cell numbers dropped to 41 and 76% of controls for primary tumor and 30 and 68% for metastases. After 72 hr exposure VP 16 had a similar effect on cells of primary tumor or metastases, but a cytotoxic effect was already evident at 0.01 microgram/ml and increased proportionally to the concentration reaching virtual complete cell killing at 1 microgram/ml. This study suggests that the much greater in vivo sensitivity to VP 16 of metastases than primary tumor of Lewis lung carcinoma probably arises less from an intrinsically greater susceptibility of metastatic cells than from pharmacokinetic factors.
Asunto(s)
Etopósido/toxicidad , Neoplasias Pulmonares/patología , Podofilotoxina/análogos & derivados , Animales , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Neoplasias Pulmonares/metabolismo , Ratones , Ratones Endogámicos C57BL , Metástasis de la Neoplasia , Neoplasias Experimentales/patología , Timidina/metabolismoRESUMEN
The in vitro cytotoxicity of 6 amino-5 formylmethylamino-1,3 dimethyluracil (ADMU) a major metabolite of caffeine in rats was studied by cell counts or [3H]thymidine incorporation in the murine Lewis lung carcinoma (3LL) and L929 fibroblast cells and in the human E cell line derived from an ovarian carcinoma. Unlike 5-fluorouracil (5FU) which was markedly cytotoxic, ADMU concentrations up to 60 micrograms/ml were devoid appreciable cytocidal action. Similarly, 1-10 micrograms 5FU markedly inhibited the blastogenic response of rat lymphocytes to PHA, whereas lymphoproliferation was not affected at ADMU concentrations up to 100 micrograms/ml. In vivo administration of ADMU (40 mg/kg, twice a day on day 1 to 3) to L1210 leukaemia-bearing mice caused a transient short-lasting reduction of tumour cell numbers only on day 6 after leukaemia inoculation.U
Asunto(s)
Cafeína/metabolismo , Uracilo/análogos & derivados , Animales , Cafeína/toxicidad , División Celular/efectos de los fármacos , Femenino , Humanos , Técnicas In Vitro , Leucemia L1210/tratamiento farmacológico , Ratones , Ratones Endogámicos , Uracilo/toxicidadRESUMEN
Thirty-two patients with by non oat cell bronchogenic carcinoma were admitted to a protocol including Cyclophosphamide (CTX) 1,000 mg/m2 i.v. day 1, VP16-213 200 mg/m2 p.o. day 1-3, every 3 weeks. Partial remissions were seen in 2 of 27 evaluable patients; 16 of 27 showed no change. Mean survival was 36.4 weeks, median survival was 38 weeks.
Asunto(s)
Carcinoma Broncogénico/tratamiento farmacológico , Ciclofosfamida/uso terapéutico , Etopósido/uso terapéutico , Neoplasias Pulmonares/tratamiento farmacológico , Podofilotoxina/análogos & derivados , Ciclofosfamida/efectos adversos , Quimioterapia Combinada , Etopósido/efectos adversos , Femenino , Humanos , MasculinoRESUMEN
It is known that human and animal fibroblasts are able to induce the retraction of a fibrin clot. In the present study the correlation between (i) fibrin-fibroblasts during growth, (ii) the number of actin stress-lines in mouse fibroblasts during growth in culture, and (iii) the sensitivity of actin stress-lines to a powerful actin-depolymerizing factor (ADF), present in plasma and serum of humans and laboratory animals was investigated. Fibroblasts at early passages (2-4) were tested for these parameters at various intervals after seeding (24, 96, and 168 h). The number of actin stress-lines was progressively higher, while the sensitivity to ADF action was progressively lower in cells cultured from 24 to 168 h; the FCR capacity was significantly decreased at 168 h. These data suggest that cells containing weakly polymerized and/or stabilized actin are more active than those containing highly polymerized and/or stabilized actin in triggering fibroblast contraction.