RESUMEN
The organization of RSV genes present in the mouse ascites sarcoma cells was analysed. The transformed cells kept in vivo and in vitro for a long time continued to contain intact RSV genome and the cellular integration site was unchanged. RSV was recovered by transfection of chick embryo cells with these cellular DNA. As these cells, especially in ascites form, grow very rapidly and it is easy to obtain a large amount of cells at a time, they are useful for further study as RSV-transformants derived from mouse cells.
Asunto(s)
Virus del Sarcoma Aviar/genética , Genes src , Genoma Viral , Sarcoma Experimental/microbiología , Integración Viral , Animales , Embrión de Pollo , ADN/aislamiento & purificación , ADN de Neoplasias/aislamiento & purificación , ADN Viral/aislamiento & purificación , Hígado/metabolismo , Ratones , Ratones Endogámicos C3H , Mapeo Restrictivo , Sarcoma Experimental/genética , Transfección , Células Tumorales CultivadasRESUMEN
PSK, a protein-bound polysaccharide derived from basidiomycetes, is a biological response modifier that exhibits a variety of activities following oral administration, including prevention of infection. In this study, the effects of oral administration of this drug on microbial flora of tumor-bearing mice were examined. Numbers of Staphylococcus, Streptococcus and Pseudomonas were increased, and those of Bifidobacterium and Lactobacillus were reduced in fresh feces of mice from later than 9 weeks after inoculation of sarcoma 180. However, these changes were prevented by oral administration of PSK. In mice bearing sarcoma 180, the increases in numbers of Staphylococcus and Pseudomonas and the decrease in those of Bifidobacterium were further enhanced by intraperitoneal administration of anticancer agent mitomycin C, but such changes were suppressed by oral administration of PSK. These results suggest that PSK has a preventive effect against abnormal conditions of the intestinal flora induced by tumor inoculation or the administration of anticancer agents.
Asunto(s)
Antibióticos Antineoplásicos/farmacología , Factores Inmunológicos/farmacología , Intestinos/efectos de los fármacos , Intestinos/microbiología , Proteoglicanos/farmacología , Sarcoma Experimental/microbiología , Administración Oral , Animales , Sinergismo Farmacológico , Heces/microbiología , Femenino , Ratones , Ratones Endogámicos ICR , Mitomicina/farmacologíaRESUMEN
The polymerase chain reaction was used for Moloney murine sarcoma virus (MoMuSV) detection in frozen and formalin-fixed, paraffin-embedded tissue sections and cultured cells isolated from MoMuSV-induced tumors. Rapid DNA extraction by proteinase K digestion, followed by CHROMA SPIN + TE-100 column purification proved to be satisfactory. Two pairs of overlapping primers, flanking 1026 base pair (bp) to 221 bp, allowed to choose among four different length of DNA-amplified segments. Although net amplification was obtained for frozen tissue and tumor cultured cells in all combinations of primers, the maximum specificity and sensitivity resulted with 602 bp fragment. This product was fully and adequately digestible using Apa I and Sau3A I restriction endonucleases. DNA extracted from paraffin-embedded sections yielded an amplification product only when the primer pair which delineated a 221-bp segment was used. This reproducible method could be useful for diagnostic and for pathogenetic investigations of MoMuSV infections.
Asunto(s)
ADN Viral/aislamiento & purificación , Virus del Sarcoma Murino de Moloney/aislamiento & purificación , Reacción en Cadena de la Polimerasa , Animales , Secuencia de Bases , Línea Celular , ADN de Cadena Simple/química , Desoxirribonucleasas de Localización Especificada Tipo II , Modelos Animales de Enfermedad , Electroforesis en Gel de Agar , Secciones por Congelación , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Virus del Sarcoma Murino de Moloney/genética , Adhesión en Parafina , Polimorfismo de Longitud del Fragmento de Restricción , Provirus/genética , Sarcoma Experimental/microbiología , Sarcoma Experimental/patología , Sensibilidad y Especificidad , Células Tumorales CultivadasRESUMEN
Primary mouse keratinocytes transformed with an activated ras oncogene transduced by helper-free Harvey sarcoma virus (HaSV) form predominantly benign tumors. In contrast, keratinocytes transformed with helper-associated HaSV form malignant tumors. We report here that this different tumorigenic behavior correlated with a much higher level of v-Ha-ras p21 protein in cells transformed with the helper-associated virus. The high level of v-ras expression in these cells was due to viral spread beyond the initial infection. The low level of v-ras p21 expression that resulted from single-hit infection with helper-free virus, together with the intrinsic heterogeneity of primary keratinocytes, explains, at least in part, the different tumorigenic behavior of keratinocytes transformed by the two types of viruses.
Asunto(s)
Transformación Celular Viral/genética , Expresión Génica/genética , Genes ras/genética , Virus del Sarcoma Murino de Harvey/fisiología , Queratinocitos/microbiología , Queratinocitos/fisiología , Proteínas Proto-Oncogénicas p21(ras)/genética , Animales , Células Cultivadas , Ratones , Sarcoma Experimental/microbiología , Sarcoma Experimental/patologíaRESUMEN
The malignant cell, in vivo, metabolizes and respires anaerobically. For a good many years scientists have been aware of anaerobia being present in malignant growths. The prevailing opinion is that the malignant cell is able to remain viable in an anaerobic medium. This assumption is incorrect and is actually misleading. My investigative studies indicates that the malignant cell metabolizes and respires intracellularly so that the consequence of such a physiology is anaerobic and the basis for such anaerobia is a metabolic phenomenon. There is a reason for this anaerobiosis. Studies demonstrated the fact that there are present within the malignant cell and in the immediate area bacterial spores arising from one of several varieties of plant bacteria. And with an adequate circulating flow of blood, by the animal (human) host, to that particular site, provides the metabolites, enzymes, and the exchange of gases to enhance this abnormal physiopathological anaerobiosis. It is this intracellular anaerobic metabolism within an animal cell that becomes the basis for its malignant transformation.
Asunto(s)
Bacterias/aislamiento & purificación , Sarcoma Experimental/metabolismo , Anaerobiosis , Animales , Bacterias/metabolismo , Bacterias/ultraestructura , Ratones , Ratones Endogámicos DBA , Microscopía Electrónica , Sarcoma Experimental/microbiología , Sarcoma Experimental/patología , Sarcoma Experimental/ultraestructura , Esporas Bacterianas/aislamiento & purificación , Esporas Bacterianas/ultraestructuraRESUMEN
Stress accelerates the growth of certain types of tumors. Here we report a possible metabolic mechanism underlying this phenomenon. Some early features of transformation include increased number of glucose transporters and greatly enhanced rates of glucose uptake; this adaptation accommodates the vast energy demands needed for neoplastic growth. In contrast, glucocorticoids, a class of steroid hormones secreted during stress, inhibit glucose transport in various tissues; this is one route by which circulating glucose concentrations are raised during stress. We reasoned that should transformed cells become resistant to this inhibitory action of glucocorticoids, such cells would gain preferential access to these elevated concentrations of glucose. In agreement with this, we observed that Fujinami sarcoma virus-transformed fibroblasts became resistant to this glucocorticoid action both in vitro and in the rat. As a result, under conditions where glucocorticoids exerted catabolic effects upon nontransformed fibroblasts (inhibition of metabolism and ATP concentrations), the opposite occurred in the virally transformed cells. We observe that this glucocorticoid resistance upon transformation cannot be explained by depletion of glucocorticoid receptors; previous studies have suggested that transformation causes an alteration in trafficking of such receptors. Because of this resistance of transformed fibroblasts to the inhibitory effects of glucocorticoids upon glucose transport, glucose stores throughout the body are, in effect, preferentially shunted to such tumors during stress.
Asunto(s)
Adenosina Trifosfato/metabolismo , Transformación Celular Neoplásica , Desoxiglucosa/metabolismo , Sarcoma Experimental/patología , Estrés Fisiológico/fisiopatología , Adrenalectomía , Animales , Transporte Biológico , Radioisótopos de Carbono , Dexametasona/farmacología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Masculino , Ratas , Ratas Sprague-Dawley , Retroviridae/genética , Retroviridae/patogenicidad , Sarcoma Experimental/metabolismo , Sarcoma Experimental/microbiologíaRESUMEN
The Schmidt-Ruppin or the B77 strain of Rous sarcoma virus (RSV) was inoculated into limb buds of 4.5-days-old avian embryos. No sarcoma but blister formation was observed in those RSV-inoculated embryos. Protein kinase activity of pp60v-src in RSV-inoculated embryos, even in the site of virus inoculation, was the same as that in mock-infected embryos. This indicated that the expression of the v-src gene did not attain superiority over that of the c-src gene in RSV-inoculated embryos. The v-src gene was detected in every DNA from tissues of RSV-inoculated embryos but not in DNAs from tissues of RSV-inoculated chicken except for the DNA from Rous sarcoma. Those results confirmed that the lack of sarcoma induction in early avian embryos by RSV was due to the lack of the expression of the v-src gene which was present in the target cells.
Asunto(s)
Virus del Sarcoma Aviar/genética , Genes src , Proteína Oncogénica pp60(v-src)/metabolismo , Proteínas Quinasas/metabolismo , Sarcoma Experimental/microbiología , Animales , Virus del Sarcoma Aviar/patogenicidad , Southern Blotting , Embrión de Pollo , ADN Viral/análisis , ADN Viral/genética , Expresión Génica , Proteína Oncogénica pp60(v-src)/aislamiento & purificación , Proteínas Quinasas/aislamiento & purificación , Sarcoma Experimental/genética , Sarcoma Experimental/patologíaRESUMEN
Rous sarcoma virus-33 (RSV-33) was obtained from a sample of chicken Rous sarcoma which had been dried and stored in 1933. RSV-33, like the RSV-29, has the minimal number of passages beyond its isolation from chicken tumour No. 1. Our experiments demonstrated that the Rous sarcoma virus-33 was replication non-defective and was pathogenic for rats. Established rat tumorigenic cell lines express the viral genome. All three species of viral RNA were detected and v-src proteins and gag polyproteins were identified as well in cells of R9 and R74 lines. The virus can be rescued from cells of R9 and R74 lines, thus indicating that the cells are virogenic. The cells of a permanent tumorigenic line RT1 are infected but not transformed by RSV-33. Although they contain a complete proviral genome, they do not express detectable virus-specific RNA. The virus is not rescuable from RT1 cells under in vivo conditions. Proviral DNA analysis showed that the RSV-33 contained a full-length genome, including the env gene, in contrast to the RSV-29 which was found replication defective.
Asunto(s)
Virus del Sarcoma Aviar/fisiología , Animales , Virus del Sarcoma Aviar/crecimiento & desarrollo , Virus del Sarcoma Aviar/patogenicidad , Línea Celular , Pollos , Cromosomas , Genoma Viral , Provirus/genética , Ratas , Ratas Endogámicas Lew , Sarcoma Experimental/química , Sarcoma Experimental/microbiología , Sarcoma Experimental/patología , Células Tumorales Cultivadas , Proteínas Virales/metabolismoRESUMEN
We have investigated the in vivo role of 2 different adhesion molecules, LFA-1 and LECAM-1, in the immune reaction to Moloney-murine-sarcoma-virus(M-MSV)-induced tumors, which undergo a peculiar spontaneous regression due to generation of a strong virus-specific cytotoxic-T-lymphocyte(CTL) response. Repeated administration of anti-LFA-1 monoclonal antibody (FD441.8 MAb), i.p. or at the site of virus inoculation, enhanced tumor growth and delayed regression, while i.p. administration of anti-LECAM-1 MEL-14 MAb gave rise to tumors that grew progressively and caused host death. Evaluation of the immunological response in MAb-treated mice showed reduced generation of virus-specific CTL precursors (p) in the spleen of animals given FD441.8 MAb i.p.; CTLp frequency in locally treated mice overlapped with that of control mice injected with virus only. FD441.8 MAb treatment did not interfere with CTL homing in the tumor, since the frequency of M-MSV-specific CTLps in sarcomas was similar in treated and control mice. Cytofluorimetric analysis indicated that the majority of tumor-infiltrating lymphocytes (TIL) from MAb-treated mice were covered by anti-LFA-1 MAb, and lacked cytotoxic activity when assayed against target cells bearing relevant tumor antigens. Instead, in mice injected i.p. with MEL-14 MAb, a very low frequency of CTLps was detected in lymph nodes draining the tumor area, and within the tumor. Our results indicate that enhanced tumor growth, depending on the MAb used, is the resultant of an inhibitory effect on different T-lymphocyte functions. Tumor progression in anti-LFA-1 MAb-injected mice is explained mostly by blockage of CTL lytic activity at the tumor site; in mice receiving i.p. MEL-14 MAb treatment, by the failure of naive T lymphocytes to enter peripheral lymph nodes and subsequently by the lack of generation of tumor-specific CTLs.
Asunto(s)
Moléculas de Adhesión Celular/inmunología , Adhesión Celular/inmunología , Antígeno-1 Asociado a Función de Linfocito/inmunología , Virus del Sarcoma Murino de Moloney/inmunología , Regresión Neoplásica Espontánea/inmunología , Sarcoma Experimental/inmunología , Linfocitos T Citotóxicos/inmunología , Animales , Anticuerpos Monoclonales , Citotoxicidad Inmunológica , Selectina L , Prueba de Cultivo Mixto de Linfocitos , Linfocitos Infiltrantes de Tumor/inmunología , Ratones , Ratones Endogámicos C57BL , Sarcoma Experimental/microbiología , Células Madre/inmunologíaRESUMEN
Previous studies have indicated that the regression versus progression of v-src-DNA-induced sarcomas is dependent on chicken line. As a first step in analyzing the role of tumor immunity as a determinant of this line dependence, experiments were undertaken to ascertain whether an antisarcoma immune response is generated in the course of sarcoma growth in TK chickens, a regressor line. To assay for this response, test TK chickens in which v-src-induced wing web sarcomas had regressed, as well as control TK chickens that had not been exposed to v-src, were challenged in protocols known to yield v-src-dependent sarcoma formation and monitored for challenge sarcoma growth. Compared with the control chickens, the test chickens showed a significant resistance to the sarcomagenic challenge. These results raise the possibility that the antisarcoma response that is inducible in regressor lines, as demonstrated here in terms of a protective effect against a subsequent sarcomagenic challenge, may also underlie the regression of v-src-induced primary sarcomas.
Asunto(s)
Virus del Sarcoma Aviar/genética , Genes src , Regresión Neoplásica Espontánea/genética , Sarcoma Experimental/microbiología , Animales , Virus del Sarcoma Aviar/fisiología , Pollos , ADN de Neoplasias/aislamiento & purificación , ADN Viral/genética , ADN Viral/aislamiento & purificación , Virus Defectuosos/genética , Virus Defectuosos/fisiología , Regresión Neoplásica Espontánea/inmunología , Regresión Neoplásica Espontánea/patología , Plásmidos , Mapeo Restrictivo , Sarcoma Experimental/inmunología , Sarcoma Experimental/patología , Especificidad de la Especie , Replicación Viral/genéticaRESUMEN
The v-erbB oncogene isolated from the R (or ES4) strain of avian erythroblastosis virus is capable of inducing erythroleukemia and fibrosarcomas. This oncogene differs from the proto-oncogene c-erbB, the avian homolog of the epidermal growth factor receptor, by its lack of an intact ligand-binding domain as well as additional alterations in its cytoplasmic coding sequences. By contrast, the insertionally activated c-erbB, a variant oncogene, which encodes a product that also lacks the ligand-binding domain but is otherwise unaltered in its cytoplasmic coding sequences, is capable of inducing leukemia but cannot induce sarcomas. In this report, we show that the critical changes for activating the sarcomagenic potential displayed by v-erbB R are two point mutations within the tyrosine kinase domain and an internal deletion of 21 amino acids in the carboxyl-terminal regulatory domain. The removal of the carboxyl-terminal autophosphorylation sites is not obligatory. These activating mutations (Arg-263 to His, Ile-384 to Ser, and the deletion of residues 494 to 514), when introduced singly into the insertionally activated c-erbB, all dramatically increase fibroblast-transforming potential. Arg-263 resides near the highly conserved HRD motif of the kinase domain, and its mutation to His increases the autophosphorylation activity. The other two mutations do not alter the intrinsic kinase activity and presumably affect other aspects of the receptor involved in growth signaling. Therefore, the high transforming potential of v-erbB R is a consequence of synergism among multiple activating mutations.
Asunto(s)
Alpharetrovirus/genética , Transformación Celular Viral , Mutagénesis Sitio-Dirigida , Proteínas Oncogénicas Virales/genética , Oncogenes , Proteínas Oncogénicas de Retroviridae/genética , Sarcoma Experimental/microbiología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Células Cultivadas , Embrión de Pollo , Humanos , Immunoblotting , Datos de Secuencia Molecular , Oligodesoxirribonucleótidos , Proteínas Oncogénicas v-erbB , Plásmidos , Proteínas Tirosina Quinasas/genética , Proteínas Tirosina Quinasas/metabolismo , Proto-Oncogenes Mas , Proteínas Recombinantes/aislamiento & purificación , Proteínas Oncogénicas de Retroviridae/aislamiento & purificación , Proteínas Oncogénicas de Retroviridae/metabolismo , Sarcoma Experimental/genética , Homología de Secuencia de Ácido Nucleico , TransfecciónRESUMEN
Herpesvirus saimiri is an oncogenic herpesvirus that induces rapidly progressing lymphomas in New World primates. Using retrovirus vectors for gene transfer, specific open reading frames of H. saimiri were tested for their ability to transform rodent cells in culture. One open reading frame, designated STP-C488 (for saimiri-transformation-associated protein of the subgroup C strain 488), phenotypically transformed Rat-1 cells, resulting in formation of foci, growth at reduced serum concentration, and growth to higher cell densities. Cells transformed by STP-C488 formed invasive tumors in nude mice. The STP-A11 reading frame of strain 11 (subgroup A) was much less potent in its transforming ability than STP-C488. These results demonstrate the oncogene nature of these two open reading frames and provide a means for studying their transforming functions independent of the rest of the H. saimiri genome.
Asunto(s)
Transformación Celular Neoplásica/genética , Genes Virales , Herpesvirus Saimiriino 2/genética , Animales , Northern Blotting , Línea Celular , ADN Viral/genética , Fibrosarcoma/genética , Fibrosarcoma/microbiología , Fibrosarcoma/patología , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Sistemas de Lectura Abierta , Plásmidos , Reacción en Cadena de la Polimerasa , Ratas , Recombinación Genética , Mapeo Restrictivo , Sarcoma Experimental/genética , Sarcoma Experimental/microbiología , Sarcoma Experimental/patología , TransfecciónRESUMEN
There are bacteria present within the malignant cell, but they are present only as spores. Although their origin stems from a branching or budding plant type of microorganism, the latter is not physically present and is not involved, per se, in the malignant cell's etiology. It is the primitive but now the asexual reproductive, conidial unicellular, ovular, or spherical shaped bodies, arising from the adult plant microorganism, in consequence to a duressing environment, but also the ones with the genetic capability of surviving within a sac or cell, that are present within the malignant cell. It is these spores, reproducing as spores within the malignant cell and within the immediate surrounding area, as long as there exists an adequate circulating flow of blood by the animal (human) host, to account for the anaerobic or preferably 'deoxygenating' metabolism that occurs in the malignant cell and is the reason for the cellular accumulation and tumor growth. This type of metabolism, respiration, and reproductive pattern is similar to that of the plant type of microorganism activity. In comparison, the healthy non-tumor tissue of animal (human) cells are genetically and primarily aerobic in respiration and metabolism. There are substantial suggestive findings and structural facts uncovered experimentally to definitely establish credence to the presence of those spores within the malignant cell and to align their presence to the pathway of activity, resulting in the etiology and physiopathology of the malignant cell and growth.
Asunto(s)
Sarcoma Experimental/fisiopatología , Esporas Bacterianas/aislamiento & purificación , Animales , División Celular , Humanos , Ratones , Ratones Endogámicos DBA , Sarcoma Experimental/microbiología , Sarcoma Experimental/patología , Esporas Bacterianas/citología , Células Tumorales CultivadasRESUMEN
Myeloproliferative syndrome was induced in adult DBA/2 mice by inoculation with myeloproliferative sarcoma virus (MPSV) and Friend murine leukemia virus (F-MuLV) as a helper virus. On day 26 after infection, the spleen weighed a maximum of 2.0 g (about 30 times the control weight). Assay of multipotent stem cells in vitro showed that the more enlarged spleens contained an increased number and concentration of mixed colony-forming units (CFU-mix) (at maximum, 11 times higher than the control). When the supernatant of cultured spleen cells was added to a serum-free bone marrow cell culture with or without erythropoietin (Epo) for detection of burst-promoting activity (BPA), it enhanced erythroid mixed colony (E-mix) formation only in the presence of Epo (p less than 0.05). Even when addition of Epo was delayed, it still induced a significant number of E-mix (p less than 0.05). These findings rule out a mimic effect of Epo resembling BPA and indicate the presence of BPA in the spleen. The culture supernatant also supported the proliferation of interleukin 3 (IL-3)-dependent 32Dcl cells. Therefore, although purification of the BPA substance has not yet been accomplished, BPA in the supernatant seems to depend on the presence of IL-3, which is known to be one of the factors stimulating multipotent hemopoietic stem cells. The presence of BPA- or CFU-mix-stimulating activity in the spleen after infection might be responsible for the development of panmyelosis, which is a characteristic of MPSV-induced myeloproliferative syndrome.
Asunto(s)
Células Madre Hematopoyéticas/patología , Interleucina-3/metabolismo , Virus del Sarcoma Murino de Moloney/fisiología , Sarcoma Experimental/patología , Neoplasias del Bazo/patología , Animales , Cromatografía por Intercambio Iónico , Ensayo de Unidades Formadoras de Colonias , Células Precursoras Eritroides/metabolismo , Células Precursoras Eritroides/microbiología , Células Precursoras Eritroides/patología , Eritropoyetina/farmacología , Femenino , Variación Genética , Hematopoyesis , Células Madre Hematopoyéticas/metabolismo , Células Madre Hematopoyéticas/microbiología , Ratones , Virus del Sarcoma Murino de Moloney/genética , Virus del Sarcoma Murino de Moloney/aislamiento & purificación , Proteínas Recombinantes/farmacología , Sarcoma Experimental/metabolismo , Sarcoma Experimental/microbiología , Neoplasias del Bazo/metabolismo , Neoplasias del Bazo/microbiología , Células Tumorales Cultivadas/metabolismo , Células Tumorales Cultivadas/microbiología , Células Tumorales Cultivadas/patologíaRESUMEN
Long-term culturing of viral Kirsten-ras-oncogene (v-Ki-ras)-transformed BALB/c 3T3 cells (KiMSV) selectively enriches for cells which have deleted the viral oncogene. In contrast, v-Ki-ras in vivo is amplified and expression increased in all late-stage tumors and lung metastases relative to late-passage KiMSV cells being injected. The nature and significance of these selection processes, in terms of the v-Ki-ras gene, have been explored using genetically-tagged cells, as have the properties of v-Ki-ras- revertant subclones. Inoculation of KiMSV late-passage cells (containing less than 5% v-Ki-ras+ cells) into nude mice, generated primary and lung metastatic tumors with the v-Ki-ras gene at increased dosage in all tumors and their single-cell clones, isolated at both early and late stages of tumor development; this demonstrates early and specific in vivo selection for v-Ki-ras+ cells in both induction and progression of tumors in this system. v-Ki-ras- revertant subclones, isolated from late-passage KiMSV cells and inoculated individually into athymic nude mice, yielded tumors for only 1 of the 4 revertants, with no evidence for v-Ki-ras sequences in these tumor cells, thereby revealing a v-Ki-ras-independent mechanism for tumor formation in a small subset of revertant cells. Mixtures of the 4 subclones yielded tumors in all animals, although at a much longer latent period than observed with v-Ki-ras+ cells. Experiments with mixtures of v-Ki-ras- revertant cells and pSV2neo0tagged/v-Ki-ras+ cells (both complex NeoR cell mixtures and individual NeoR clones tested) at various cell ratios revealed clonal variability among v-Ki-ras+ cells for dominance during tumor formation. Moreover, the complex NeoR cell mixtures yielded both primary and metastatic tumors with simplified patterns of pSV2neo integration sites, suggesting that secondary genetic or epigenetic events, in addition to v-Ki-ras, contribute to the tumor-progressing phenotype. These experiments taken together demonstrate (a) clone-specific early selection of distinct v-Ki-ras+ cells amongst themselves and over v-Ki-ras- cells in both induction and progression of tumors, (b) reduced tumorigenic competence of v-Ki-ras- revertant cells, with a small subset displaying a v-Ki-ras-independent mechanism for tumor formation in this BALB/c 3T3 system, and (c) the significance of additional genetic or epigenetic events for tumor-progressing competence in unique subsets of v-Ki-ras+ cells.
Asunto(s)
Transformación Celular Viral , Genes ras , Virus del Sarcoma Murino de Kirsten/genética , Sarcoma Experimental/patología , Animales , Southern Blotting , Células Clonales , Sondas de ADN , ADN de Neoplasias/genética , ADN de Neoplasias/aislamiento & purificación , ADN Viral/genética , ADN Viral/aislamiento & purificación , Genes Dominantes , Cinética , Ratones , Ratones Endogámicos BALB C , Sarcoma Experimental/microbiología , TransfecciónRESUMEN
The long terminal repeats (LTR) of the defective murine sarcoma viruses (MSV) containing v-abl, v-Ha-ras, or v-fos were exchanged for LTRs from other retroviruses having different tissue tropism. The new chimeric MSV were found to induce the same diseases as the parental viruses, indicating that sequences outside the LTR, most likely those of the oncogene, are responsible for the disease specificity of these defective MSV.
Asunto(s)
Transformación Celular Neoplásica , Virus Defectuosos/genética , Genes abl , Genes ras , Proteínas Proto-Oncogénicas/genética , Proto-Oncogenes , Secuencias Repetitivas de Ácidos Nucleicos , Virus del Sarcoma Murino/genética , Sarcoma Experimental/microbiología , Animales , Línea Celular , Quimera , Virus Defectuosos/patogenicidad , Genes Virales , Ratones , Ratones Endogámicos , Proteínas Tirosina Quinasas/genética , Proteínas Proto-Oncogénicas c-fos , Virus del Sarcoma Murino/patogenicidadRESUMEN
Previous results have established that subcutaneous inoculation of chickens (line SC) with a v-src(+) subviral DNA fragment induces the formation of progressor sarcomas at the wing web site of inoculation. Because the sarcoma cells are incompetent for production of exogenous progeny virus, this system is a useful model of tumor expansion by sarcoma cell division, in the absence of infection-mediated recruitment of new tumor cells. The present study was undertaken to define conditions that modulate the pattern of growth (regression vs progression) of v-src DNA-induced sarcomas. These conditions were found to include the line of chicken or the presence on the subviral v-src(+) DNA fragment of a viral replication-specific sequence that includes env.
Asunto(s)
Virus del Sarcoma Aviar/genética , Genes src , Sarcoma Experimental/microbiología , Animales , Virus del Sarcoma Aviar/patogenicidad , Pollos , ADN Viral/genética , Plásmidos , Mapeo Restrictivo , Sarcoma Experimental/patologíaRESUMEN
We investigated DNA damage caused by carcinogenic metals in a murine sarcoma virus (MuSV)-based mutagenicity assay in which mutations targeted to v-mos expression can be selected. Nickel chloride treatment of NRK cells (termed 6m2 cells) infected with MuSVts110, a retrovirus conditionally defective in viral RNA splicing and cell transformation, caused the outgrowth of transformed "revertants" with changes in the MuSVts110 RNA splicing phenotype. Cadmium and chromium treatment of 6m2 cells resulted in the selection of a second class of revertants with what appeared to be frameshift mutations allowing the translation of a readthrough gag-mos protein. In both classes of metal-induced revertants, viral gene expression was distinct from that observed in revertants arising in untreated 6m2 cultures, arguing that metal treatment did not simply enhance the rate of spontaneous reversion. In one representative nickel revertant line the operative nickel-induced mutation affecting MuSVts110 RNA splicing was a duplication of 70 bases surrounding the 3' splice site. The effect of this mutation was to direct splicing to the most downstream of the duplicated 3' sites and concomitantly relax its characteristic thermosensitivity. These data establish the mutagenic potential of nickel and provide the first example of a defined nickel-induced mutation in a mammalian gene.
Asunto(s)
Daño del ADN , ADN Viral/efectos de los fármacos , Regulación Viral de la Expresión Génica/efectos de los fármacos , Níquel/toxicidad , Virus del Sarcoma Murino/genética , Sarcoma Experimental/genética , Animales , Cadmio/toxicidad , Cromo/toxicidad , Deleción Cromosómica , Pruebas de Mutagenicidad , Fenotipo , Sarcoma Experimental/microbiología , Temperatura , TransfecciónRESUMEN
N-tropic murine leukemia viruses have been observed in connection with radiation-induced osteosarcomagenesis in BALB/c mice. We have investigated the bone disease-inducing potential of molecularly cloned, BALB/c-derived N-tropic viruses in the random-bred NMRI mouse strain. The germ-line virus and an exogenous virus isolate were found to induce high incidences of osteopetrosis and lymphomas and a lower incidence of osteomas. Two viruses derived from somatically acquired proviruses of independent radiation-induced osteosarcomas induced lower incidence of osteopetrosis and lymphomas. Nucleotide sequence analysis of the long terminal repeat regions and RNase T1 fingerprint analysis revealed only few differences between the isolates. The possible involvement of N-tropic murine leukemia viruses in radiation-induced osteosarcomagenesis in the BALB/c mouse strain is discussed.
Asunto(s)
Virus de la Leucemia Murina/patogenicidad , Ratones Endogámicos BALB C/microbiología , Animales , Secuencia de Bases , Clonación Molecular , Virus de la Leucemia Murina/genética , Ratones , Mapeo Nucleótido , Osteosarcoma/microbiología , ARN Viral/química , Sarcoma Experimental/microbiologíaRESUMEN
Permanent alterations of the epithelial differentiation pattern were investigated after infection of HH-16 cl. 4 adenocarcinoma cells with Moloney murine sarcoma virus (MoMuSV). Transformed cell clones with fibroblastoid morphology were isolated and compared with clones of unchanged epithelioid phenotype. Southern blot analyses showed intact MoMuSV proviral genomes in copy numbers between 4 and 9 in the DNA of the morphologically transformed cell clones as well as in the clones with unaltered morphology. The fibroblastoid cells produced sarcomas after inoculation of newborn rats, whereas MoMuSV-infected cell clones with unaltered epithelioid morphology yielded adenocarcinomas. Immunocytochemical analyses revealed that morphological transformation into the fibroblastoid phenotype was accompanied by loss of cytokeratin expression and appearance of the mesenchymal marker protein vimentin. Proviral DNA was transcribed in the infected cell clones irrespective of their phenotype; however, transcription was significantly higher in cells with fibroblastoid morphology than in epithelioid cells.