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PubMed1998

Novel p53 tumour suppressor mutations in cases of spindle cell sarcoma, pleomorphic sarcoma and fibrosarcoma in cats.

Twenty feline neoplasms were sequenced in the region from exons 5 to 8 for the presence of tumour suppressor gene p53 mutations. In a spindle cell sarcoma of the bladder, a missense mutation (codon 164 AAG-->GAG, lysine-->glutamic acid) in exon 5 was detected. In a pleomorphic sarcoma, a 23 bp deletion involving the splicing junction between intron 5 and exon 6 was observed. In a fibrosarcoma, a 6 bp deletion of p53 covering 2 bp of exon 7 and 4 bp of intron 7, including the splicing junction, was found. The study demonstrates three new p53 mutations in different types of sarcomas in cats.

Amino Acid Sequence↗

PubMed2002

Cumulative influence of elastin peptides and plasminogen on matrix metalloproteinase activation and type I collagen invasion by HT-1080 fibrosarcoma cells.

HT-1080 fibrosarcoma cells express at their plasma membrane the elastin-binding protein (EBP). Occupancy of EBP by elastin fragments, tropoelastin or XGVAPG peptides was found to trigger procollagenase-1 (proMMP-1) overproduction by HT-1080 cells at the protein and enzyme levels. RT-PCR analysis indicated that elastin peptides did not modify the MMP-1 mRNA steady state levels, suggesting the involvement of a post-transcriptional mechanism. We previously reported that binding of elastin peptides to EBP induced other matrix metalloproteinases (MMP-2 and MT1-MMP) expression. Since those peptides were here found to also accelerate the secretion of urokinase from HT-1080 cells, culture medium was supplemented with plasminogen together with elastin peptides at aims to induce or potentiate MMPs activation cascades. In such conditions, plasmin activity was generated and exacerbate proMMP-1 and proMMP-2 activation. As a consequence, elastin peptides and plasminogen-treated HT-1080 cells displayed a significant type I collagen matrix invasive capacity.

Collagen Type I↗

PubMed1972

The effect of cyclophosphamide on the growth and cellular kinetics of a transplantable rat fibrosarcoma.

The gross response of a transplantable fibrosarcoma (RIB(5)) in rats treated with a single dose of 10, 50, 100 or 200 mg/kg body weight of cyclophosphamide (CP) is reported. The variation in mitotic and tritiated thymidine ((3)H-TdR) labelling indices after a single dose of 100 mg/kg CP was studied. Examination of chromosome spreads has shown the time course of the visible damage to DNA caused by CP and a transient arrest of cells in G(2) was demonstrated by microdensitometry. The effects of 100 mg/kg body weight CP given in 2 equal doses separated by various time intervals were examined in an attempt to relate tumour response to its altered cellular state at the time of the second dose.

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PubMed1973

A quantitative morphological analysis of the response of a transplantable rat fibrosarcoma to cyclophosphamide.

The response of a transplantable rat fibrosarcoma, RIB(5), to a single injection of 100 mg/kg body weight of cyclophosphamide, 0·5, 1, 2 and 3 days after treatment, has been studied by means of a morphometric analysis of tumour sections and has been compared to the changes occurring in untreated tumours. The results showed a transitory increase in thymidine labelling index 12 hours after drug injection accompanied by a low mitotic index which persisted for a further 12 hours. These observations are compared with, and confirm, a previous study involving analysis of smear preparations from tumour cell suspensions. The changes in the proportion of the tumour occupied by erythrocytes and necrotic elements are described. The results of the morphometric analysis have shown the changes occurring in the outer, intermediate and inner zones of the tumour. In all control tumours, and tumours up to 2 days after cyclophosphamide treatment, necrosis was predominant in the inner zone and tumour cell proliferation was most marked in the outer zone. In contrast, 3 days after cyclophosphamide treatment proliferation was predominant in the inner zone and the amount of necrosis was approximately the same in each zone.

Animals↗

PubMed1975

Effect of local injection of Corynebacterium parvum on the growth of a murine fibrosarcoma.

Local injection (i.e. injection at the site of tumour inoculation) of strains of C. Parvum which have a significant anti-tumour effect when given systemically (i.e. intravenously or, in the case of subcutaneous tumour transplant, intraperitoneally) strongly inhibits the growth of isogeneic transplants of a fibrosarcoma in intact CBA mice but has little or no effect on subcutaneous transplants of the same tumour in T-cell deprived mice. The anti-tumour effect of local injection of C. parvum, unlike that of systemic injection in this particular tumour system, thus appears to be T-cell dependent.

Animals↗

PubMed1976

Studies on the Fc receptor bearing cells in a transplanted methylcholanthrene induced mouse fibrosarcoma.

The presence of Fc receptors on the surface of cell suspensions obtained from a transplanted isogeneic methylcholanthrene induced murine fibrosarcoma has been investigated by determining the capacity of such cells to form rosettes with antibody coated SRBC. These studies indicate that a large percentage of cells in the tumour had Fc receptors on their surface. The proportion of such cells was increased by reducing the number of cells transplanted, by administering cyclophosphamide to the host, and on occasions by the i.p. injection of C. parvum. It was largely unaffected by the route of tumour cell transplantation or by T cell depletion of the host before transplantation but appeared to decline in older (i.e. larger) tumours. Both phagocytic and non-phagocytic cells had Fc receptors on their surface. The phagocytic population appeared to be affected most by procedures which altered the overall percentage of Fc receptor bearing cells. The Fc receptor bearing tumour cells were separated from those devoid of Fc receptors on the basis of their adherent properties. Upon transplantation to isogeneic hosts both populations gave rise to tumours containing a high percentage of Fc receptor bearing cells. These studies suggest that many of the Fc receptor bearing cells in our tumour are probably infiltrating cells of host origin. Their significance in relation to tumour growth remains to be established.

Animals↗

PubMed1977

Non-specific cytotoxicity of spleen cells in mice bearing transplanted chemically induced fibrosarcomas.

Spleen cells collected from mice bearing transplanted chemically induced syngeneic fibrosarcomas non-specifically inhibited DNA synthesis of sarcoma and lymphoma target cells in vitro. Splenocytes from mice hyper-immunized against a syngeneic sarcoma specifically inhibited DNA synthesis of the tumour used for immunization. The impairment of tumour-cell DNA synthesis was associated in vitro with cytostasis, and lysis of the target cells was not seen. Since treatment with anti-theta serum and complement did not impair cytostatic action of the spleen cells, and since thymus-deprived animals showed similar activity to normal mice, T lymphocytes were not involved in non-specific cytostasis. Removal of phagocytic adherent cells by carbonyl iron markedly inhibited the cytostatic activity of the spleen cells, suggesting a role in this reaction for cells of the monocyte-macrophage series. The presence of an actively growing sarcoma was a prerequisite for the expression of non-specific cytostasis, since surgical excision resulted in complete disappearance of this activity of spleen cells.

Animals↗

PubMed1977

Effect of X-irradiation on host-cell infiltration and growth of a murine fibrosarcoma.

Whole body X-irradiation (400 rad) of C57BL mice, either before or after i.m. implantation of the syngeneic fibrosarcoma, FS6, influenced both the growth of the tumours and their cellular composition, particularly their macrophage content. Pre-irradiation resulted in slower initial growth of tumours, and a concomitant lack of host-cell infiltration, but when tumours began to grow at a rate parallel to controls infiltration by host cells was demonstrable. Similarly, irradiation of the tumour-bearing host resulted in a temporary cessation of growth, and a decrease in the macrophage content, which did not return to control levels for 2-3 weeks after irradiation. The significance of these results is discussed in relation to the possibility that infiltrating host cells, particularly macrophages, may stimulate the growth of this tumour.

Animals↗

PubMed1978

Lack of correlation between in vivo rejection of syngeneic fibrosarcomas and in vitro non-specific macrophage cytotoxicity.

Two transplantable, highly immunogenic syngeneic C57BL fibrosarcomas, FS1 and FS6, were shown to have tumour-specific rejection antigens, as shown by excision of the primary tumours and i.p. or i.m. injection of graded doses of the specific and unrelated tumour cells. I.p. challenge with tumour cells induced a large and relatively long-lasting increase in numbers of peritoneal leucocytes. Macrophage monolayers prepared from such exudates were, in general, non-specifically cytotoxic, though occasional specific cytotoxicity was detected. T lymphocytes isolated from exudates were shown to kill in a specific manner. When immunized mice were challenged with the specific tumour cells to elicit large numbers of peritoneal cytotoxic cells, and with graded doses of the non-cross-reacting tumour cells at the same time or at various times thereafter, growth of the non-related tumours occurred in all cases and only the specific tumour was rejected. Moreover, Winn tests, in which the inflammatory cells were mixed with unrelated tumour cells and implanted i.m., did not delay tumour growth. The relevance of these findings to the role of macrophages and lymphocytes in syngeneic tumour rejection is discussed.

Animals↗

PubMed1979

Phase-specific cytotoxicity in vivo of hydroxyurea on murine fibrosarcoma cells synchronized by centrifugal elutriation.

The S-phase-specific cytotoxicity of hydroxyurea (HU) was tested on synchronized murine fibrosarcoma (FSa) cells lodged in the lungs of C3Hf/Bu mice. FSa cells from primary asynchronous cultures were separated and synchronized on the basis of size by centrifugal elutriation. Flow microfluorometry (FMF) was used to determine the cell-cycle parameters and the relative synchrony of the separated populations. After elutriation, 8000 viable FSa cells from each fraction, along with 10(6) heavily irradiated tumour cells (unseparated) were injected i.v. into whole-body-irradiated mice (20 per group). Under these conditions, 95% of the injected cells, regardless of size or position in the cell cycle, are arrested in the lungs. Twenty minutes later, hydroxyurea (HU, 1 mg/g) was administered i.p. into 10 animals of each group. Fourteen days later the animals were killed, their lungs removed and fixed, and the number of macroscopic tumour nodules counted. Killing of the initially injected cells by HU, as evidenced by a reduction in lung colonies in treated animals, correlated with the precentage of S-phase cells in each fraction. The greatest effect, an 80% reduction in colony number, was seen in Fraction 8, containing the largest percentage of S-phase cells (65%). These results demonstrate the usefulness of this procedure as a rapid method for characterizing the phase specificity of chemotherapeutic drugs in vivo.

Animals↗

PubMed1980

Cytotoxic effect in vivo of selected chemotherapeutic agents on synchronized murine fibrosarcoma cells.

The cytotoxic effects in vivo of single doses of either adriamycin (ADM), 1-beta-D-arabinofuranosylcytosine (Ara-C), bleomycin (BLM), cis-diamminedichloroplatinum (II) (cis-DDP), or cyclophosphamide (CY) on murine fibrosarcoma (FSa) cell populations were determined. Tumour cells were separated and synchronized by centrifugal elutriation. Viable tumour cells from selected elutriator fractions were then injected i.v. into whole-body-irradiated mice. Twenty minutes later selected doses of ADM, Ara-C, BLM, cis-DDP or CY were administered to selected groups of these animals. Fourteen days later the mice were killed. Killing of injected tumour cells by each of the chemotherapeutic agents was evidenced by a reduction in the lung cells by each of the chemotherapeutic agents was evidenced by a reduction in the lung colonies per cell injected in treated animals. Under these conditions the response of FSa cells in vivo to the 5 drugs tested differed both qualitatively and quantitatively. Ara-C was S-phase-specific in toxicity. ADM, BLM, and cis-DDP were preferentially toxic to S, G2+M and G1 cells respectively. CY, a drug requiring bioactivation to form alkylating metabolites, was found to be equally toxic to G1 and G2+M enriched populations, but less effective in killing cell populations enriched with early-S cells.

Animals↗

PubMed1980

Malignant properties and DNA content of daughter clones from a mouse fibrosarcoma: differentiation between malignant properties.

Freshly isolated cones of high cloning efficiency from a mouse fibrosarcoma were examined for DNA content, cell size, protein content, and malignant characteristics such as artificial lung-colony-forming ability, s.c. tumour take, host survival, and spontaneous metastatic ability. These malignant characteristics and other cell properties were heterogeneous among these clones; the malignant characteristics could vary and were not "all or none" in their nature. The higher the DNA content or the larger the cell volume, the higher the malignancy in terms of artificial lung-colony forming, efficiency, s.c. tumour take, and host survival. Despite variability of each parameter, the ratio of DNA content to cell size or protein content remained constant through these variations: the increased DNA paralleled increased protein and increased cell volume. The increased DNA was correlated with the more malignant characteristics of local growth and lung-colony-forming efficiency. Spontaneous metastasis to the lung was totally different from the local growth abilities; the small-cell clone produced more metastases. The graded nature of malignant properties and the differentiation between local growth and metastatic potential among the daughter clones indicate that malignancy reflects a complex moiety of cell properties.

Animals↗

PubMed1981

Promotion of fibrosarcoma cell growth by products of syngeneic host macrophages.

Cells from a C57BL/cbi chemically induced fibrosarcoma (FS6) require exogenous platelet-derived growth factor (PDGF) for in vitro proliferation (as do normal "untransformed" fibroblasts) whereas cells obtained from the FS6M1 tumour, a spontaneous metastasizing subline, show autonomy from PDGF in vitro. Furthermore, the FS6 cells exhibit very low colony formation in an anchorage-independent growth assay. In vivo, this tumour is immunogenic, rarely metastasizes and is heavily infiltrated by host macrophages. Studies of in vitro cell proliferation and anchorage-independent growth show that syngeneic host macrophages from the peritoneal cavity or from the growing tumour release a diffusible factor(s) which has (1) growth-stimulating activity on FS6 cells in monolayer cultures in PDGF-poor medium and (2) potent colony-stimulating activity on FS6 cell cultured in methyl-cellulose-containing medium. These macrophage supernatants stimulate proliferation of quiescent normal fibroblasts in monolayer culture as well as FS6 sarcoma cells, but do not stimulate anchorage-independent growth of normal cells. Supernatants from BCG-elicited macrophages were shown to contain abundant arginase, and were cytolytic to FS6 cells but not to normal cells. Heat inactivation abrogated the arginase and cytotoxicity, revealing heat-stable mitogenicity for FS6 cells and normal fibroblasts. The stimulatory effect of macrophages on FS6 sarcoma cells can be mimicked by the addition of the tumour promoter 12-tetradecanoyl-phorbol-13-acetate (TPA) and supports the hypothesis that macrophages could play a significant role in multistage carcinogenesis by providing a source of endogenous promoter.

Animals↗

PubMed1982

Decrease in tumour growth by injections of histamine or serotonin in fibrosarcoma-bearing mice: influence of H1 and H2 histamine receptors.

C3H and C57 BL/6 mice carrying methylcholanthrene-induced fibrosarcomas were injected i.p. daily with histamine, metiamide (anti-histamine type-2 receptor), histamine + metiamide, mepyramine (anti-histamine type-1 receptor), serotonin and methysergide (anti-serotonin). Inhibition of tumour growth and lengthened survival were observed with histamine and histamine + metiamide. The best results (both on tumour growth and survival) were obtained with serotonin. Survival was increased by metiamide and decreased by mepyramine and methysergide. In histamine-treated and in serotonin-treated mice, histological studies of the tumours showed large and numerous foci of haemorrhagic necrosis. Stimulation of histamine type-1 or serotonin receptors and inhibition of histamine type-2 receptors play a beneficial role in the host's defence against tumours.

Animals↗

PubMed1982

Phase-specific cytotoxicity in vivo of hydroxyurea on murine fibrosarcoma pulmonary nodules.

The cytotoxic effects in vivo of hydroxyurea (HU) on murine fibrosarcoma (FSa) cells grown as pulmonary tumours were determined. Tumour cells from 13-day-old nodules were made into suspension and separated on the basis of cell size by centrifugal elutriation. Flow microfluorometry (FMF) was used to determine the cell-cycle parameters and the relative synchrony of the separated populations, as well as the degree of contamination by normal diploid cells in each of the tumour-cell populations. HU cytotoxicity was tested by administering both a single 1 mg/g i.p. dose into mice that had been injected i.v. 20 min earlier with known numbers of synchronized viable FSa cells, and i.p. doses of 1 mg/g each into mice bearing 13-day-old pulmonary nodules. In the latter experiments, animals were killed 1 h after the last dose, and the tumour nodules were excised and made into a single-cell suspension and elutriated. Known numbers of cells from each fraction were injected into recipient mice to determine survival. In both sets of experiments, cell killing by HU correlated with the percentage of S-phase cells. The treatment of 13-day-old pulmonary nodules with 3 doses of HU also depleted the (G2+M) phase tumour cells and increased the heterogeneity between tumour subpopulations, as determined by FMF analysis.

Animals↗

PubMed1985

The effect of passage in vitro and in vivo on the properties of murine fibrosarcomas. II. Sensitivity to cell-mediated cytotoxicity in vitro.

The sensitivity of cultured and mouse-passaged cloned lines of chemically-induced murine fibrosarcomas to killing by NK and NC cells, and to cell-mediated immunity, has been studied in in vitro assays, using target cells labelled with 51Cr or 125IUDR. None of the lines tested proved sensitive to NK cells. Three cultured lines were, at most, only slightly sensitive to NC cells; a fourth cultured line was moderately sensitive and became less so, but not completely insensitive, after passage in susceptible hosts. The primary object of these experiments was to test the hypothesis that cultured cell lines which ordinarily fail to grow in normal mice are able to grow after being passaged in a susceptible immunodeficient host because, during this passage, they become resistant to NK or NC cells. This has been shown to occur with one clone, but will not serve as a general explanation because, with other clones, both cultured and mouse-passaged lines were NC-insensitive. The cell-mediated immunity assays confirm our previous conclusion that cultured and mouse-passaged lines of the same clone differ little, if it all, in immunogenicity.

Animals↗

PubMed1985

The effect of passage in vitro and in vivo on the properties of murine fibrosarcomas I. Tumorigenicity and immunogenicity.

Cloned cell lines of chemically-induced murine fibrosarcomas maintained in tissue culture usually fail to grow when transplanted to normal syngeneic mice. They grow, however, in various categories of T cell deficient mice and after such passage grow readily in normal mice. Both cultured and mouse-passaged lines possess strong TATA. Three alternative explanations are suggested which might account for these findings. Emergence during the initial passage of a population of tumour cells resistant to NC cells. Acquisition during the initial passage of a protective surface molecule that interferes with the efferent side of the immune response when the tumour cells are subsequently transplanted to a normal host. Loss during the initial passage of a Class I MHC molecule which prevents dual recognition of the tumour cells by T cells when they are transplanted to a normal host. New experiments are proposed to distinguish between these possibilities.

Animals↗

PubMed1985

In vivo detection and partial characterization of effector and suppressor cell populations in spleens of mice with large metastatic fibrosarcomas.

The MC-2 fibrosarcoma, which is a transplantable tumour syngeneic for BALB/c mice, metastasizes to lymph nodes draining subcutaneous inoculation sites, and also to the lungs. T cell-mediated immunity was detected in Winn assays using spleens from excision immunized mice. T cell-mediated anti-tumour immunity was also detected in spleens from mice with small tumours but disappeared as the tumour burden increased. Protective immune spleen cell activity in the Winn assay was inhibited by prior addition of spleen cells from mice with large tumours, causing increased tumour incidence. Splenic metastases occasionally occurred in the MC-2 model, but were not demonstrable by bioassay in any of the experiments detecting splenic suppressor cell activity. In vivo protective activity was restored to advanced-stage tumour-bearer spleens by whole-body ionizing irradiation (0.5 and 2.5 Gy) of donor mice 15 h before sampling. Spleen cells from mice with small tumours remained protective after 1.5, 2.5 and 4.0 Gy of irradiation in vivo. These results are consistent with the properties of radiosensitive suppressor T cells. In contrast to reports in other tumour models, suppressor cells were not detected until late in the course of MC-2 development. This is surprising in view of the aggressively metastatic nature of MC-2. It is postulated that modulation of the anti-tumour immune response by the suppressor cells is associated with metastasis in this tumour model. The late appearance of both suppressor cells and metastatic cells in the spleen may reflect similar processes occurring earlier in regional lymph nodes.

Animals↗