RESUMEN
Three to 20 hr after an i.p. injection of 64Cu (half-life, 12.8 hr) into mice bearing Krebs ascites cells, a high amount of the radioisotope was recovered in the ascites cells themselves. In the control group, the radioisotope was mainly present in the liver. Similar amounts of 64Cu were recovered in regenerating as well as in normal liver, whereas in the liver of mice bearing ascites cells, this amount was lower by 40 to 50% regardless of the ascitic volume. Thus, the copper metabolism seems to be disturbed at the hepatic level in mice bearing ascites cells. The distribution of 64Cu was 'analyzed in DNA, RNA, and proteins from cellular lysates fractionated by CsCl gradient. There was a uniform pattern of distribution in the macromolecules from ascites cells, while 64Cu' was preferentially associated with the protein fraction from liver. Further experiments indicated that, in vivo, 64Cu was bound to the DNA of ascites cells.
Asunto(s)
Carcinoma Krebs 2/metabolismo , Cobre/metabolismo , ADN/metabolismo , Animales , Líquido Ascítico/análisis , Fraccionamiento Celular , Cobre/análisis , ADN/análisis , Femenino , Inyecciones Intraperitoneales , Hígado/metabolismo , Ratones , Proteínas/análisis , ARN/análisis , Radioisótopos/metabolismo , Distribución TisularRESUMEN
Previous experiments have shown that 64Cu transmutation was inefficient to cure mice which had been injected 6 days earlier with 5 x 10(5) Krebs ascitic cells. The experiments were repeated with 64CuCl2 administered only 1 day after the injection of 5 x 10(5) Krebs ascitic cells when no developing tumor existed. The results reported showed that, even in this case, only a delay in death could be observed. These negative results revealed that malignant cells injected 24 hours previously in a mouse (in vivo conditions) differ from a malignant cell suspension in a tube (in vitro conditions) where the lethal effect of 64Cu transmutation was clearly evidenced. We concluded that some kind of collaboration was established between the few accepted malignant cells and the host. Based on this collaboration we introduced a new concept, "the cancer mouse", in which in addition to the malignant cells some cells in the host, even though non - malignant, nonetheless feature a slightly modified functioning: "the cancer functioning". In this view, an efficient antitumor treatment must act at the same time and in a coordinated manner on the malignant cells and on the host cells which now feature "the cancer functioning". In the efficient treatment described (Anticancer Res 9: 947-954, 1989) some compounds (64Cu and thioproline) act against the malignant genomes, and other components (metal ions, amino acids, vitamin D2, thyroxine and chelating substances) act against the functioning of the host cells by taking into account certain characteristics of living systems. This treatment was efficient in curing mice injected 1 or 6 days previously with 5 x 10(5) Krebs ascitic cells.
Asunto(s)
Carcinoma Krebs 2/radioterapia , Radioisótopos de Cobre/uso terapéutico , Modelos Teóricos , Animales , Carcinoma Krebs 2/fisiopatología , Relación Dosis-Respuesta en la Radiación , Femenino , Ratones , Factores de TiempoRESUMEN
The treatment described previously (Anticancer Res 9:947-954,1989) was efficient when applied on day 1 or 6 after the injection of 5 x 10(5) Krebs ascitic cells in mice. Under our experimental conditions, all the untreated mice died within 12 to 25 days. The experiments described show that the treatment developed, is efficient when applied on day 11 but not on day 16. To understand the difference in the treatment efficiency between these two days, we tested the 64Cu incorporation inside the ascitic cells. It was observed that 64Cu incorporation exists on day 11 but no longer on day 16. On day 16, the inefficiency of the treatment must be correlated with the non-incorporation of 64Cu inside the ascitic cells. As the tumor growth is also arrested on day 16, an irreversible stage is reached. A model was developed to explain the results obtained. In this model, cancer develops in 3 successive stages. In the first stage, the cellular functioning is under the control of the malignant tumor cells and the number of cells with the "cancer functioning" is increasing with time, but decreasing after removal of the malignant tumor; in the second stage, tumor and cancer both develop independently, meaning that the number of cells with the "cancer functioning" will continue to increase after the removal of the malignant tumor; in the third stage, each cell of the organism has the "cancer functioning" and the characteristics of malignancy will by retroaction.
Asunto(s)
Carcinoma Krebs 2/radioterapia , Radioisótopos de Cobre/uso terapéutico , Modelos Teóricos , Animales , Carcinoma Krebs 2/patología , Cobre/análisis , Femenino , Ratones , Estadificación de Neoplasias , Radioterapia/métodos , Factores de TiempoRESUMEN
A new treatment is proposed for mice bearing a Krebs ascitic tumor targeting on both the malignant cells and the host cells having the "cancer functioning" (cancer cells) (Anticancer Res 9: 941-946 1989). It is shown that the treatment (a) injure specifically via 64Cu transmutation the DNA of the malignant cells and further perform (with thioproline or spermine) a "reverse transformation" on the damage DNA; (b) restore a "noncancer functioning" in the host cells which had become "cancer cells"; this restoration was performed using, at physiological concentrations, natural compounds already present in all cell types such as metal ions, amino acids, vitamin D2, thyroxine and chelating substances. To decrease the damage induced in the organism by the radiation emitted by 64Cu, a radioprotector, glycerol, was used. Two different strains of mice were used (Swiss or CBA). For each of them, the same treatment was efficient on condition that thioproline or spermine was used for Swiss or CBA mice respectively.
Asunto(s)
Carcinoma Krebs 2/radioterapia , Radioisótopos de Cobre/uso terapéutico , Modelos Teóricos , Animales , Carcinoma Krebs 2/fisiopatología , Daño del ADN , Femenino , Ratones , Ratones Endogámicos CBA , Radioterapia/métodos , Espermina/uso terapéutico , Factores de TiempoRESUMEN
In the efficient treatment described (Anticancer Research 9: 941-946, 1989) some compounds (64Cu and thioproline) act against the malignant genomes, and other components (metal ions, amino acids, vitamin D2, thyroxine and chelating substances) act against the functioning of the host cells having the cancer functioning (cancer cells). The experiments described show that this treatment developed with mice injected on day zero with 5 x 10(5) Krebs ascitic cells, is also efficient for mice bearing a spontaneous malignant tumor even if it is not an ascitic tumor. After the decrease in weight due to the loss of the tumor, a typical weight evolution was observed characterized by a second weight increase followed by a decrease leading to normal weight. This last weight remained stable. This typical weight evolution in tumor-free mice (after spontaneous tumor or experimental Krebs ascitic tumor) was related to the age of the tumor at the beginning of the treatment, that is, according to our hypothesis (Anticancer Res 9:955-960, 1989) to the cancer evolution emphasizing the great importance of all physiological phenomena related to this cancer course. To test the consequence of this treatment, some of these tumor-free mice were injected with 5 x 10(5) ascitic cells. No tumor developed in these tumor-free mice meaning that they also were tumor-resistant. This last result is in agreement with the forecast of the systemic analysis on which the efficient treatment was based.
Asunto(s)
Radioisótopos de Cobre/uso terapéutico , Neoplasias Experimentales/fisiopatología , Animales , Secuencia de Bases , Peso Corporal , ADN de Neoplasias/análisis , Femenino , Ratones , Ratones Endogámicos CBA , Trasplante de Neoplasias , Neoplasias Experimentales/radioterapiaRESUMEN
65-Zinc distribution was studied in different organs of either healthy mice or mice bearing an ascitic tumor. This study clearly showed that zinc distribution differs between healthy and tumoral mice. Moreover, zinc distribution in the different organs of the mice is modified by the quantity administered.
Asunto(s)
Ascitis/metabolismo , Neoplasias Experimentales/metabolismo , Zinc/farmacocinética , Animales , Relación Dosis-Respuesta a Droga , Ratones , Distribución Tisular , Zinc/administración & dosificación , Radioisótopos de ZincRESUMEN
64Cu was injected in the form of CuCl2 either by subcutaneous or by intraperitoneal route, and its distribution inside different organs was analyzed in 5 different tumor models, 4 in mice and 1 in rats. In all organs tested (blood, liver, kidneys, spleen, intestine, muscle, and tumor) no significant differences were observed in the results obtained after either injection route. All tumors analyzed (Krebs ascite, intestinal Leiomyo sarcoma, human tumor, mammary adenocarcinoma, either spontaneous or chemically induced) contained a relatively high concentration of 64Cu. For all tumor models tested, the 64Cu distribution was altered as compared with that of the corresponding control animals.
Asunto(s)
Cobre/metabolismo , Neoplasias Experimentales/metabolismo , Adenocarcinoma/metabolismo , Animales , Femenino , Masculino , Neoplasias Mamarias Experimentales/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones Desnudos , Radioisótopos , Ratas , Ratas Endogámicas F344 , Distribución TisularRESUMEN
The decays of 64Cu incorporated in human malignant (A549) or monkey nonmalignant (CVI) cells lead to cell death. When plotted as a function of the radioactivity introduced in the growth medium (microCi/ml at t = 0), the residual colony-forming capability decreases exponentially. The slope of the corresponding curve is steeper for A549 than for CV1 cells. Different data show that the cellular lethal event is a consequence of 64Cu transmutation and not of the irradiation by the simultaneously emitted beta- and beta+ particles. Liquid holding results show that the lethal event is irreparable. The decays of 67Cu, another radioisotope of copper, lead to cell death with the same exponential survival curve and the same lethal efficiency as for 64Cu, in spite of their different decay schemes. The lethal efficiency of both copper isotopes is close to that of 125I utilized in the form of iododeoxyuridine under the same experimental conditions as 64Cu and 67Cu. The high lethal efficiency of radioactive copper transmutations raises questions about the role in DNA functioning of copper atoms known to be present in trace amounts in this macromolecule. The lethal consequence of radioactive copper transmutations suggests that the copper atoms bound to DNA are essential for cellular functioning.
Asunto(s)
Supervivencia Celular/efectos de la radiación , Radioisótopos de Cobre , Animales , Línea Celular , Haplorrinos , Humanos , Radiactividad , Células Tumorales Cultivadas/efectos de la radiaciónRESUMEN
We have recently demonstrated the existence of a lethal effect due to the transmutation of 64Cu atoms associated with the DNA of in vitro cultured cells: normal monkey cells (kidney) and malignant human cells (alveolar lung carcinoma). The lethal efficiency per decay is high and even higher for the malignant cells. From these results, we have tried to bring about a regression of an ascitic tumor developing in the mouse. The experiments we described show that it is possible to clearly delay (5 X 10(5) ascitic cells injected at t = 0), even to stop for a given number of animals (1 to 2 X 10(4) ascitic cells injected at t = 0), the growth of the tumor owing to some injections of 64Cu totalling about 3 mCi and carried out from the 6th day following inoculation of the malignant cells.
Asunto(s)
Carcinoma Krebs 2/radioterapia , Cobre/uso terapéutico , Radioisótopos/uso terapéutico , Animales , Ascitis , ADN de Neoplasias/biosíntesis , Femenino , RatonesRESUMEN
CV1 and A549 cells were grown in the presence of 64Cu porphyric complex, 64CuCl2, or 67CuCl2. Radioactive copper determinations were performed on whole cells and on isolated cellular DNA. 125IUdR was used to calibrate the particular extraction and purification procedures we developed because of the half-lives of 64Cu and 67Cu. The results obtained have shown that some radioactive copper atoms remained firmly bound to the DNA molecule. Their amount was of the same order when using two different DNA isolation methods for the two cell lines studied. No significant differences were found when 64Cu was used as CuCl2 or as porphyric complex.
Asunto(s)
Cobre/metabolismo , Idoxuridina/metabolismo , Animales , Línea Celular , Células Cultivadas , Centrifugación por Gradiente de Densidad , Radioisótopos de Cobre , ADN/aislamiento & purificación , Etanol , Semivida , Haplorrinos , Humanos , Radioisótopos de Yodo , Riñón/citología , TripsinaAsunto(s)
Isótopos de Carbono/análisis , Pronasa , Tritio/análisis , Animales , Femenino , Masculino , Métodos , Ratones , Radiactividad , CintigrafíaAsunto(s)
Neoplasias Mamarias Experimentales/metabolismo , Agua/metabolismo , Tejido Adiposo/metabolismo , Glándulas Suprarrenales/metabolismo , Animales , Femenino , Semivida , Glándulas Mamarias Animales/metabolismo , Tasa de Depuración Metabólica , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Ratones Endogámicos , Músculos/metabolismo , Ovario/metabolismo , Hipófisis/metabolismo , Tritio/sangreAsunto(s)
Acetato de Clormadinona/metabolismo , Neoplasias Mamarias Experimentales/metabolismo , Tejido Adiposo/metabolismo , Glándulas Suprarrenales/metabolismo , Animales , Cruzamientos Genéticos , Femenino , Semivida , Intubación Gastrointestinal , Glándulas Mamarias Animales/metabolismo , Tasa de Depuración Metabólica , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos ICR , Ratones Endogámicos , Músculos/metabolismo , Hipófisis/metabolismo , Factores de Tiempo , TritioRESUMEN
The experiments show the lethal effect of the beta decay of 33P incorporated in DNA of bacteriophage S 13. The lethal efficiency is high, 0.72 at 0 degrees C and 0.55 at--197 degrees C. The presence of a radical scavenger like AET has no influence. It was found previously that for such phages with single-stranded DNA, the lethal efficiency of 32P decay is unity, and that the lethal event is a DNA single-strand break, owing to the high energy of the nucleogenic 32S atom. As the recoil energy of the 33S atom is too low to account for such a break, it is suggested that the reorganization of the phosphate molecule into sulphate is able to bring about a DNA single-strand break with an efficiency as high as 0.7, at 0 degrees C. A model for the DNA double-strand-break produced by a transmutation processes is suggested.
Asunto(s)
Bacteriófagos/efectos de la radiación , ADN Viral/efectos de la radiación , Radioisótopos de Fósforo , Bacteriófagos/metabolismo , ADN de Cadena Simple/efectos de la radiación , Electrones , Escherichia coli , Modelos Biológicos , Fosfatos/metabolismo , Fosfatos/efectos de la radiación , Radiación Ionizante , Radiactividad , Sulfatos/metabolismo , Sulfatos/efectos de la radiación , TemperaturaRESUMEN
To study the lethal effect of 64Cu under in vitro conditions, asynchronous mammalian cells were used. A lethal effect does exist as a consequence of the decay itself of a few 64Cu atoms incorporated in cellular DNA. This lethal effect is characterized by an exponential survival curve with no shoulder and no tail; it exists even for non-dividing cells. The lethal efficiency per decay is very high. To test the 64Cu lethal effect under in vivo conditions, experiments were performed with ascitic cells developing in mice. In this case, the lethal effect also exists, but it is not a function of the 64Cu doses injected in the mice. Faced with this puzzling result, a systemic approach was necessary to understand and counteract ascitic cells developing in mice.
Asunto(s)
Cobre/uso terapéutico , Neoplasias Experimentales/radioterapia , Radioisótopos/uso terapéutico , Animales , Líquido Ascítico/patología , Línea Celular , Supervivencia Celular/efectos de la radiación , Células Cultivadas , Cobre/metabolismo , Daño del ADN , ADN de Neoplasias/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Ratones , Neoplasias Experimentales/patología , Radiactividad , Radioisótopos/metabolismo , Distribución TisularRESUMEN
When circular single-stranded DNA of phage S13 is labelled with 32P or 33P, the transmutations very efficiently bring about a loss of phage infectiousness (efficiency = 1 for 32P and 0.73 for 33P). For both radionuclides, the lethal efficiencies as well as the lethal events are different. In the case of 32P, the lethal event is the loss of the circular integrity of the DNA molecule, occurring as a consequence of a systematic single strand-break caused by each 32P decay (100%). Conversely, in the case of 33P, the lethal events are either a single strand-break (40%) or a local stereochemical modification (33%). The same primary event, the substitution at each 33P decay of a phosphate by a sulfate molecule, leads to one of these lethal events in relation to the decay site. Moreover, neither the phage adsorption nor its genome injection into bacteria depends on the physical state of the genome, and thus lethality is revealed at only the genetic level.
Asunto(s)
Colifagos/efectos de la radiación , Daño del ADN , ADN Viral/efectos de la radiación , Escherichia coli/efectos de la radiación , Genes Virales/efectos de la radiación , Centrifugación por Gradiente de Densidad , Colifagos/genética , ADN Viral/genética , ADN Viral/aislamiento & purificación , Escherichia coli/genética , Mutación , Radioisótopos de FósforoRESUMEN
A high lethal efficiency was observed when the decay of two radioactive isotopes of copper, 64Cu or 67Cu, occurred in mammalian cells. The lethal efficiency is of the same order for both isotopes in spite of their different decay processes. As the lethal event could be attributed to an injury inside the cellular DNA only, these results suggest that, even if present in the DNA in trace amounts solely, copper atoms are essential chromatin components. Their behaviour differs depending on whether the cell is a tumor or a non tumor cell because their lethal efficiencies are different (0.5 and 0.1 respectively).