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Pulsed DC electric fields couple to natural NAD(P)H oscillations in HT-1080 fibrosarcoma cells.

Previously, we have demonstrated that NAD(P)H levels in neutrophils and macrophages are oscillatory. We have also found that weak ultra low frequency AC or pulsed DC electric fields can resonate with, and increase the amplitude of, NAD(P)H oscillations in these cells. For these cells, increased NAD(P)H amplitudes directly signal changes in behavior in the absence of cytokines or chemotactic factors. Here, we have studied the effect of pulsed DC electric fields on HT-1080 fibrosarcoma cells. As in neutrophils and macrophages, NAD(P)H levels oscillate. We find that weak (approximately 10(-5) V/m), but properly phased DC (pulsed) electric fields, resonate with NAD(P)H oscillations in polarized and migratory, but not spherical, HT-1080 cells. In this instance, electric field resonance signals an increase in HT-1080 pericellular proteolytic activity. Electric field resonance also triggers an immediate increase in the production of reactive oxygen metabolites. Under resonance conditions, we find evidence of DNA damage in HT-1080 cells in as little as 5 minutes. Thus the ability of external electric fields to effect cell function and physiology by acting on NAD(P)H oscillations is not restricted to cells of the hematopoietic lineage, but may be a universal property of many, if not all polarized and migratory eukaryotic cells.

Biological Clocks↗

Identification of a laminin-like substance on the surface of high-malignant murine fibrosarcoma cells.

High- and low-malignant murine fibrosarcoma cells were stained with anti-laminin antibodies using immunoperoxidase techniques and examined by electron microscopy. With the high-malignant cells, specific staining was observed along the cell surface. Use of normal rabbit serum in place of the rabbit anti-laminin or pretreatment of the anti-laminin with soluble laminin completely eliminated this staining. No immunoperoxidase staining was observed with the low-malignant cells. In additional studies, membrane fractions were prepared from the high- and low-malignant cells and used to immunize rabbits. The animals immunized with the membrane fractions from the high-malignant cells produced antibodies that reacted by enzyme-linked immunosorbent assay (ELISA) with murine laminin obtained from the EHS sarcoma. The animals immunized with membrane fractions from the low-malignant cells did not. These studies provide strong evidence that the high-malignant cells (but not the low) express on their cell surface a substance that is immunologically cross-reactive with laminin. In addition, the high-malignant cells (but not the low) secreted a material into the cell culture fluid that could be specifically immunoprecipitated with antilaminin antibodies. The immunoprecipitated material co-migrated with purified laminin when examined by sodium dodecyl sulphate/polyacrylamide gel electrophoresis under reducing conditions. The existence of this substance associated with the surface of the high-malignant cells and its absence from that of the low-malignant cells may explain the previously noted difference between these cells in their ability to attach to type IV collagen. This difference may also contribute to the dissimilarity between these cells in their metastatic potential.

Animals↗

Quiescence and increased adhesion after mammalian DNA transfection of human fibrosarcoma cells.

When a population of human fibrosarcoma HT1080 cells was transfected with a mammalian expression vector and DNA pieces representing either the human whole genome or mouse bulk cDNA, there was a transient increase in the number of adhesive cells in the population. The number of cells with increased adhesion was proportional to the amount of transfected DNA; the increase occurred at a maximal frequency of between 10(-4) and 10(-5) per cell. Increased adhesion in these cells persisted for 9-12 days, corresponding to the period of highly efficient transient transfection, and was accompanied by arrest in cell division. Transfection of non-mammalian DNAs, reduction of transfected mammalian sequence length by restriction enzyme digestion, or omission of expression vector DNA did not permit these shifts in phenotype. The effects seen suggest that expression of specific transfected mammalian DNA sequences suppresses certain phenotypic characteristics in these transformed mammalian cells.

Cell Adhesion↗

Chemo-immunotherapy of methylchoranthrene-induced fibrosarcoma by concanavalin A-bound tumor vaccine, levamisole and mitomycin C.

Concanavalin A (ConA)-bound-tumor cell vaccine of methylchoranthrene-induced fibrosarcoma (Meth 1) induced tumor-specific immunoprophylactic and immunotherapeutic response against an inoculum of live Meth 1 cells in histocompatible animals. ConA-free Meth 1 vaccine induced much less response under the identical experimental conditions. Immunotherapeutic potency of ConA-bound Meth 1 vaccine was enhanced by levamisole, and 37% of the animals inoculated with 10(4) live Meth 1 cells at day 0 were cured when they were administered 10(6) cells of ConA-bound Meth 1 vaccine at days 1 and 8 and 0.63 mg/kg levamisole at days 1,2 and 3. Delayed administration of levamisole at day 8,9 and 10 was less effective than the earlier administration, but still produced a 17% cure of Meth 1-bearing animals when combined with ConA-bound Meth 1 vaccine. Immunotherapeutic response under these regimens was further enhanced by mitomycin C, and approximately 60% of the animals inoculated with 10(5) Meth 1 cells were cured when three agents were administered at the defined intervals. These results suggest the feasibility of the regimen in which the therapeutic response induced by immunotherapeutic agents is further enhanced by the selected chemotherapeutic agents.

Animals↗

Bradykinin-induced cyclic AMP accumulation in mouse fibrosarcoma independent of prostaglandin E2 formation.

The relationship between bradykinin (BK)-induced prostaglandin E2 (PGE2) and cyclic AMP syntheses in mouse fibrosarcoma cells (HSDM1C1) was investigated. Maximal BK-induced increases in cyclic AMP preceded increases in PGE2 production. PGE2 synthesis reached maximum at a much lower concentration of BK than cyclic AMP synthesis. Indomethacin completely inhibited BK-induced PGE2 production, but did not influence the cyclic AMP levels. Arachidonic acid in the medium induced PGE2 production in large quantities, but increased cyclic AMP accumulation only slightly. A high PGE2 concentration increased cyclic AMP levels only slightly. Theophylline increased basal and BK-mediated cyclic AMP levels, but did not affect PGE2 production at all. These results indicate that BK-evoked PGE2 and cyclic AMP syntheses in HSDM1C1 are not dependent upon each other.

Animals↗

Fibrosarcomas induced by cobalt chloride (CoCl2) in rats.

Hyperlipaemia persisted for more than 12 months in rats treated with cobalt chloride, 9 of 20 treated rats and 1 control rat died within the first year, and 8 of the surviving animals developed fibrosarcomas--in 4 of these the tumour was far removed from the injection sites. The results are discussed with reference to the use of cobalt chloride treatment for anaemia in patients with chronic renal failure.

Animals↗

RBE of neutrons generated by 50 MeV deuterons on beryllium for control of artificial pulmonary metastases of a mouse fibrosarcoma.

Artificial pulmonary metastases of a mouse fibrosarcoma were produced by the intravenous injection of 10(4) cells admixed with 2 X 10(6) plastic microspheres into mice preconditioned with 600 rad whole-body irradiation 24 hours earlier. Four days after injection of tumour cells, mice were irradiated with neutrons generated by 50 MeV deuterons on Be at the Texas A & M Variable Energy Cyclotron or with 137Cs gamma rays. One, three or six fractions of radiation were delivered on a three-hour fractionation schedule. Surviving lung metastases were scored macroscopically 16 days after irradiation. The data indicate that: (1) the RBE (n/gamma) was in the range 1.6-2.6 depending on the size of dose per fraction; (2) the slopes of the gamma-ray curves decreased with increasing fraction number (i.e. decreasing fraction size); (3) the slopes of the neutron curves decreased only slightly with increasing fraction number (and decreasing fraction size); (4) no additional sparing was achieved by further fractionating doses of neutrons of 300 rad or less.

Animals↗

Response of selected tumour cell populations separated from a fibrosarcoma following irradiation in situ with fast neutrons.

We have characterized selected populations of fibrosarcoma cells separated on density gradients of Renografin with respect to distribution in the cell cycle, clonogenic ability, and response to radiation. The latter two parameters have been observed to be related to the density of the cells, the denser populations being less clonogenic but more resistant to both low and high LET radiation. We have determined that these cells effectively repair potentially lethal damage if allowed to remain in situ following exposure to low LET radiation. This repair ability is not observed, however, following neutron irradiation. In addition, cells left in situ are relatively more sensitive to low doses of either low or high LET radiation. This difference in response is believed to be due to differences in the expression of damage by cells left in situ as compared to those assayed immediately. These data reflect the usefulness of cell separation methods in the study of solid tumour systems. Specifically, these methods reduce the heterogeneity found in the solid tumour by allowing for the isolation of subpopulations which then can be studied either individually or in relation to the entire tumour.

Animals↗

Relative biological effectiveness of 290 MeV/u carbon ions for the growth delay of a radioresistant murine fibrosarcoma.

The relative biological effectiveness (RBE) for animal tumors treated with fractionated doses of 290 MeV/u carbon ions was studied. The growth delay of NFSa fibrosarcoma in mice was investigated following various daily doses given with carbon ions or those given with cesium gamma-rays, and the RBE was determined. Animal tumors were irradiated with carbon ions of various LET (linear energy transfer) in a 6-cm SOBP (spread-out Bragg peak), and the isoeffect doses; i.e. the dose necessary to induce a tumor growth delay of 15 days were studied. The iso-effect dose for carbon ions of 14 and 20 keV/microm increased with an increase in the number of fractions up to 4 fractions. The increase in the isoeffect dose with the fraction number was small for carbon ions of 44 keV/microm, and was not observed for 74 keV/microm. The alpha and beta values of the linear-quadratic model for the radiation dose-cell survival relationship were calculated by the Fe-plot analysis method. The alpha values increased linearly with an increase in the LET, while the beta values were independent of the LET. The alpha/beta ratio was 129 +/- 10 Gy for gamma-rays, and increased with an increase in the LET, reaching 475 +/- 168 Gy for 74 keV/microm carbon ions. The RBE for carbon ions relative to Cs-137 gamma-rays increased with the LET. The RBE values for 14 and 20 keV/microm carbon ions were 1.4 and independent of the number of fractions, while those for 44 and 74 keV/microm increased from 1.8 to 2.3 and from 2.4 to 3.0, respectively, when the number of fractions increased from 1 to 4. Increasing the number of fractions further from 4 to 6 was not associated with an increase in the RBE. These results together with our earlier study on the skin reaction support the use of an RBE of 3.0 in clinical trials of 80 keV/microm carbon beams. The RBE values for low doses of carbon beams were also considered.

Animals↗

Radiosensitizer sanazole (AK-2123) enhances gamma-radiation-induced apoptosis in murine fibrosarcoma.

Sanazole (AK-2123) (N-2'-methoxy ethyl)-2-(3"-nitro-1"-triazolyl)acetamide, which has completed phase III clinical trials as a radiosensitizer, enhanced gamma-radiation induced apoptosis in murine fibrosarcoma upon i.p. administration at 40 mg/kg body weight one hour prior to irradiation. A microscopic examination of Giemsa-May-Grunwald stained cells has shown a higher frequency of condensed nuclei and fragmented nuclei in the tumor cells. The administration of sanazole to tumor-bearing animals enhanced the radiation-induced internucleosomal fragmentation in the nuclear genome of tumor cells. Higher levels of caspase-3 activity were also observed in the cell extracts of tumours from AK-2123 administered mice. Exposure to gamma-radiation of AK-2123-treated mouse further enhanced the caspase-3 activity, indicating the induction of apoptosis. The radiation sensitization property of sanazole was discernible by comparing the relative tumor diameter following irradiation after i.p. administration of AK-2123 and irradiation alone; it was higher during the first few days followed by the treatment.

Animals↗

Fibrosarcoma of the head and neck in children.

A retrospective study of 11 children with fibrosarcoma of the head and neck is presented. Six of these tumors are well differentiated, and three are poorly differentiated. Nine patients were followed for over six years and are available for treatment evaluation. The results of treatment indicate that patients with superficially located, well differentiated tumors occurring at sites where wide surgical margins can be obtained have the most favorable prognosis.

Child↗

Fibrosarcoma of the head and neck.

A review of 118 archival cases originally diagnosed as fibrosarcoma of the head and neck is presented. Sites of involvement were the sinonasal region, upper aerodigestive tract, skin, and soft tissues. Review of the pathologic specimens applying strict histopathologic criteria including immunohistochemical techniques reclassified many of the tumors as malignant fibrous histiocytoma, monophasic synovial sarcoma-like neoplasm, or dermatofibrosarcoma protuberans. Improved clinical outcome correlated significantly with lesser tumor size, lower histologic grade, and negative surgical margins. Location in the skin or soft tissue tended to convey improved survival, although not to a level of statistical significance.

Adult↗

Interactions of asbestos-activated macrophages with an experimental fibrosarcoma.

Supernatants from in vivo asbestos-activated macrophages failed to show any cytostatic activity against a syngeneic fibrosarcoma cell line in vitro. UICC chrysotile-induced peritoneal exudate cells also failed to demonstrate any growth inhibitory effect on the same cells in Winn assays of tumor growth. Mixing UICC crocidolite with inoculated tumor cells resulted in a dose-dependent inhibition of tumor growth; this could, however, be explained by a direct cytostatic effect on the tumor cells of high doses of crocidolite, which was observed in vitro.

5'-Nucleotidase↗

Activation of anchorage-independent growth of HT1080 human fibrosarcoma cells by dexamethasone.

Anchorage independence is an important hallmark of the transformation that correlates with tumorigenicity. We have isolated a variant clone of HT1080 human fibrosarcoma cells (cl-2) that is specifically defective in anchorage-independent growth. Interestingly, 10(-7) M dexamethasone (DEX) substantially rescued the anchorage-independent growth of cl-2 cells in semisolid culture. DEX also promoted the anchorage-independent growth of parental HT1080 cells. However, the agent had no effect on the anchorage-dependent growth of cl-2 and parental cells in ordinary liquid culture. Cell cycle analysis demonstrated that the population of G0/G1 cells increased, whereas that of S and G2/M cells decreased in growth-arrested cl-2 cells in suspension culture. However, such an effect of anchorage loss on cell cycle progression was alleviated by adding 10(-7) M DEX. In cl-2 cells in semisolid culture, DEX suppressed the expression of P27Kip1, whereas it stimulated the expression of cyclin A and hyperphosphorylated retinoblastoma (Rb) proteins. On the other hand, DEX had no effect on cyclin D1 and P21Cap1 expression. These effects of DEX, except for the suppression of P27Kip1, were blocked by an antimicrofilament drug, cytochalasin D. Our results suggest that the stimulation of anchorage-independent growth by DEX involves at least two regulatory mechanisms, i.e., one that leads to the suppression of P27Kip1 protein without requiring cytoskeletal integrity, and another that requires cytoskeletal integrity, leading to stimulation of cyclin A and hyperphosphorylation of Rb protein.

Base Sequence↗

Primary cardiac fibrosarcoma in a dog.

A primary cardiac fibrosarcoma in the right atrium of a 6-year-old Chihuahua dog is described. At necropsy, there was a firm, whitish and spherical mass in the right atrium. Histopathologically, the mass had moderate cellularity composed of spindle-shaped cells with scattered multinucleated giant cells. The tumor cells were arranged in interwoven bundles and sheets in the collagenous stroma. No metastases were observed. Ultrastructurally, the tumor cells mainly consisted of fibroblasts. Multinucleated giant cells did not have any certain organelles that would indicate a higher order of differentiation. Primary cardiac sarcomas in dogs are extremely rare.

Animals↗

Congenital-infantile fibrosarcoma. A clinicopathologic study of 10 cases and molecular detection of the ETV6-NTRK3 fusion transcripts using paraffin-embedded tissues.

Congenital-infantile fibrosarcoma (CIFS) is a relatively indolent sarcoma that should be distinguished from more aggressive spindle cell sarcomas of childhood. CIFSs have been found to have a novel recurrent reciprocal translocation t(12;15)(p13;q25) resulting in the gene fusion ETV6-NTRK3 (ETS variant gene 6; neurotrophic tyrosine kinase receptor type 3). We studied immunohistochemical expression of NTRK3, and conducted a reverse transcription-polymerase chain reaction (RT-PCR) assay to detect the ETV6-NTRK3 fusion transcripts using archival formalin-fixed paraffin-embedded tissues from 10 CIFSs. Thirty-eight other spindle cell tumors were included as controls. The ETV6-NTRK3 fusion transcripts were identified in 7 (70%) of 10 CIFSs. Nucleotide sequence analysis showed that the fusion occurred between ETV6 exon 5 and NTRK3 exon 13. The 38 control tumors were negative for the fusion transcript. Immunohistochemically, CIFSs consistently expressed NTRK3. But the expression of NTRK3 also was observed in 22 of 38 control tumors. These results show the diagnostic usefulness of RT-PCR methods to detect ETV6-NTRK3 fusion transcripts in archival formalin-fixed paraffin-embedded tissue and the important role of NTRK3 in the development of CIFS, despite its being a protein of little importance in differential diagnosis.

Child↗

Keloidal fibromas and fibrosarcomas in the dog.

Sixteen dogs (2-12 years of age) presented with one (n = 15) or two (n = 1) cutaneous nodules (n = 16) or a dermal plaque (n = 1). Intact males (n = 9) and neutered males (n = 4) were more affected than were females (n = 3). Histologically, these lesions were characterized by focal dermal and subcutaneous deposition of thick hyalinized collagen fibers intermingled with fibroblasts, and in 13 of 17 lesions, a variable number of CD18-positive cells were interpreted as reactive macrophages. Fibroblasts in three dogs formed intersecting fascicles, interpreted as evidence of malignant transformation. The terms keloidal fibroma and keloidal fibrosarcoma can be applied to these lesions. Excision was curative in five dogs with keloidal fibroma for which follow-up was available. However, because malignant transformation may occur, wide excision of canine keloidal lesions is warranted.

Animals↗

Fibrosarcoma of the mandible: a case report.

Fibrosarcoma is a relatively rare malignant tumor found in the head and neck region. Pain and swelling are the two most frequent symptoms, but other findings include paresthesia, trismus, loosening of teeth, ulceration, swelling, and pathological fracture. This case report underscores the importance of early diagnosis and detection of oral neoplasms to minimize therapeutic intervention.

Adult↗