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[Demonstration of SYT-SSX11/2 fusion transcripts in synovial sarcomas using RT-PCR].

Various mesenchymal malignancies with a spindle cell morphology may mimic monophasic synovial sarcomas. We analysed, whether detection of SYT-SSX1/2 fusion transcripts, present as a consequence of a specific translocation [t(X;18)(p11.2;q11.2)] found in synovial sarcomas, are useful to confirm the diagnosis of synovial sarcoma. A nested RT-PCR protocol was established for the detection of SYT-SSX1/2 fusion transcripts. After RNA extraction of snap frozen tissue reverse transcription was carried out. In a nested PCR, the resulting cDNA samples were amplified and visualized on agarose gels. DNA sequencing of PCR products enabled the assignment of fusion transcripts to either the SYT-SSX1 or the SYT-SSX2 variant. We detected SYT-SSX1/2 fusion transcripts in seven monophasic and three biphasic synovial sarcomas. 20 out of 21 control tumour including leiomyosarcomas, malignant peripheral nerve sheath tumours, gastrointestinal stromal sarcomas and fibrosarcomas were negative in RT-PCR analysis. One case of recurrent spindle cell sarcoma originally classified as a fibrosarcoma revealed a SYT-SSX2 fusion transcript. These data provide further evidence that the RT-PCR amplification of SYT-SSX1/2 fusion transcripts permits the specific identification of synovial sarcomas. This diagnostic approach may be especially useful in cases with equivocal histomorphology.

Chromosome Mapping↗

[Histological aspects of the sarcomatous degeneration of bone in Paget's disease].

After a critical review of the literature and an enquiry based on 70 cases, the author confirms that osteogenic sarcomas and fibrosarcomas constitute the usual means of sarcomatous degeneration in bones affected by Paget's disease. Giant-cell tumours present a distinct group. The problems posed by the initial stages of the malignant transformation and the osteosarcomas with multiple centres are then discussed.

Bone Neoplasms↗

The tumor rejection antigen separated from Rous sarcoma virus-induced murine fibrosarcoma exhibits a molecular weight of approximately 60 kD but differs from functional pp60src.

The tumor antigen capable of inducing tumor resistance (tumor rejection antigen; TRA) was obtained in a solubilized form by sodium dodecyl sulfate (SDS) extraction of plasma membrane fraction from Rous sarcoma virus (RSV)-induced CSA1M fibrosarcoma cells (BALB/c origin). Analyses by Sephacryl S-300 gel filtration and SDS-polyacrylamide gel electrophoresis revealed that TRA activity was recovered in the fraction with a molecular weight of approximately 60 kD. Unfractionated crude SDS-solubilized preparation contained gp70 as detected by rabbit anti-gp70 antiserum, whereas such reactivity was lost in the fraction exhibiting the molecular weight of about 60 kD. Since this fraction retained pp60src activity, the relation of TRA to pp60src was further investigated. pp60v-src was also obtained from the lysate of v-src-expressing yeast transformant. Immunization of BALB/c mice with such pp60v-src-containing lysate failed to induce any significant tumor protection. The above 60 kD fraction of CSA1M solubilized antigens was allowed to bind to Sepharose beads coupled with anti-pp60src monoclonal antibody and separated into the bead-bound and bead-unbound fractions. The bead-bound fraction that was recovered from pp60src-binding beads (pp60src-positive fraction) did not exhibit the TRA activity. In contrast, immunization with the fraction depleted of pp60src activity (bead-unbound fraction) resulted in potent tumor protection. These results indicate that the solubilized membranous component(s) of CSA1M with a molecular weight of approximately 60 kD, which is distinct from functional pp60src, functions as the TRA against RSV-induced CSA1M tumor cells.

Animals↗

Nonangiogenic and nonlymphomatous sarcomas of the canine spleen: 57 cases (1975-1987).

The case records of and histopathologic findings in 57 dogs with nonangiogenic and nonlymphomatous splenic sarcomas were reviewed. Splenic neoplasms in these dogs included leiomyosarcoma, fibrosarcoma, undifferentiated sarcoma, liposarcoma, osteosarcoma, chondrosarcoma, myxosarcoma, rhabdomyosarcoma, and fibrous histiocytoma. The clinical signs associated with splenic sarcoma included anorexia or decreased appetite, abdominal distention, polydipsia, lethargy, vomiting, weight loss, and weakness. An abdominal mass was detected in 86% of the dogs by use of abdominal palpation (63%), and/or abdominal radiography (74%). The diagnosis was based on histopathologic findings in the spleen. Abdominal exploratory surgery was performed on 43 of the 57 dogs. Twenty-seven dogs were treated by splenectomy, and 16 were euthanatized at the time of surgery because of widespread metastatic lesions. Of the 14 dogs on which surgery was not performed, 11 were euthanatized on the basis of results of preoperative diagnostic tests, and the remaining 3 dogs had splenic neoplasms that were incidental findings at necropsy. Of the 27 surgically treated dogs, 5 died in the immediate postoperative period, 12 died or were euthanatized within 1 year after splenectomy, and only 5 dogs survived greater than or equal to 1 year. Three dogs were lost to follow-up evaluation, and 2 were still alive 6 and 7 months after surgery. The median survival time of the 22 dogs for which survival was known was 2.5 months. The median survival time for 11 dogs with no obvious metastasis at the time of splenectomy was 9 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Pathology of soft tissue sarcomas: 238 cases of the childhood tumor registry].

Until April 1981 malignant soft tissue sarcomas were registered from 238 patients. Rhabdomyosarcoma was the most common tumor (115/238 = 48.3%). The embryonal subtype was predominantly seen among the rhabdomyosarcomas (83/115 = 72.2%). Rhabdomyosarcomas were localized most frequently in the head and neck area (40/115 = 34.8%), followed by genitourinary system (15/115), pelvis soft tissue (12), abdomen (10) and extremities (10). Non-rhabdomyosarcomatous soft tissue sarcomas (123/238 = 51.7%) were synovial sarcomas (20 = 8.4%), fibrosarcomas including spindle cell sarcoma (17 = 7.4%), leiomyosarcomas (12 = 5.0%), malignant tumors of the vascular system (11 = 4.6%) and neurofibrosarcomas (9 = 3.8%). Other types of sarcoma were extremely rare. 42 (17.6%) of all soft tissue sarcomas could not be classified histogenetically. Rhabdomyosarcomas could be diagnosed much more accurately (105/115 = 91.3%), compared to all other soft tissue sarcomas (99/121 = 81.8%). At present, the most difficult diagnostic problems remain with the tumors of connective tissue, in particular with fibrosarcomas and with the differential diagnosis of juvenile fibrosarcomas versus juvenile fibromatoses.

Adolescent↗

Primary thoracic sarcomas.

Primary sarcomas of the thorax are rare. The diagnosis is established only after sarcomalike primary lung malignancies and metastatic disease have been excluded. Primary sarcomas of the thorax are classified according to their histologic features and constitute a large group of tumors that occur in the lung, mediastinum, pleura, and chest wall. Angiosarcoma, leiomyosarcoma, rhabdomyosarcoma, and mesothelioma (sarcomatoid variant) are the most common primary intrathoracic sarcomas. Ewing sarcoma, primitive neuroectodermal tumor, chondrosarcoma, malignant fibrous histiocytoma, osteosarcoma, synovial sarcoma, and fibrosarcoma usually arise in the chest wall. Although primary thoracic sarcomas commonly manifest as large, heterogeneous masses, they have a wide spectrum of radiologic manifestations, including solitary pulmonary nodules, central endobronchial tumors, and intraluminal masses within the pulmonary arteries. The different histologic types of sarcomas are frequently indistinguishable at radiologic analysis. However, differences in clinical presentation and the location of the tumor, as well as morphologic features such as calcification within the mass and rib involvement, can be useful in suggesting the appropriate diagnosis. For example, a large rib mass in a child with fever and malaise indicates a Ewing sarcoma, a mass with a calcified matrix is likely a chondrosarcoma or osteosarcoma, and a pulmonary artery mass is likely a leiomyosarcoma.

Adult↗

An infallible method for producing ascitic fluid from solid tumours.

Notwithstanding all precautions, the methods usually applied for the transformation of solid tumours into the ascitic form, frequently fail. This must certainly be ascribed to the still existing, considerably high intercellular adhesion forces despite the fact that cancer cells generally show lack of adhesiveness due to their higher electrical charge in comparison with normal cells. Hagmar recently stipulated that particularly anionic detergents increase the negative surface charge of tumour cells, while cationic detergents drastically reduce or even reverse this charge. A series of comparative experiments were undertaken by us in order to study the influence of anionic, nonionic and amphoteric detergents on the production of ascitic fluid from macerates of Yoshida sarcoma and fibrosarcoma BUSP. This latter tumour was produced and maintained in our laboratory and resisted to all our attempts to produce the ascitic form. Heparin, in concentrations of 0,2, 0,4 and 1,0 mg/ml, was used as an example of an anionic detergent; Pluronic F-68 and phosphatidylcholine, both in concentrations of 1, 2, and 5 mg/ml, represent a nonionic and an amphoteric detergent, respectively. The obtained results clearly show a most favourable action of the applied surfactants: whereas the controls, in which the detergents were not applied, failed to produce ascitic fluid in, sometimes, considerable percentages (up to 100% in the case of BUSP) of the treated animals, not a single animal remained free of ascite when the injected macerate was pre-treated with the emulsifier. It should here be mentioned, however, that heparin in the highest concentration applied (1 mg/ml) caused lysis of the tumour cells impeding, thus, whatever formations of ascitic fluid.

Animals↗

Suicide gene therapy of sarcoma cell lines using recombinant adeno-associated virus 2 vectors.

Soft-tissue sarcomas are mesenchymal tumors that respond poorly to systemic chemotherapy. Suicide gene therapy may be an alternative treatment strategy. Here we show a high susceptibility of human sarcoma cell lines for recombinant adeno-associated virus 2 (rAAV-2) suicide vectors: connective tissue sarcoma (HS-1), fibrosarcoma (HT-1080), Ewing sarcoma (RD-ES), Askin tumor (SK-N-MC), rhabdomyosarcoma (A-204) and soft-tissue sarcoma (WSKL-1). Several vectors containing the thymidine kinase (TK) gene under the control of either the cytomegalovirus promoter or the elongation-factor 1 alpha (EF1alpha) promoter were cloned and tested. Higher expression levels of the transgene were observed in the sarcoma lines when using the EF1alpha-suicide gene-containing vectors. A complete eradication of rAAV-2-EF1alpha-TK/eGFP (TK/enhanced green fluorescent protein fusion gene)-transduced tumor cells was shown following exposure to ganciclovir (2.5 microg/ml) in vitro, while at this dose level > 90% of mock-transduced tumor cells survived. Xenotransplantation tumor models (intraperitoneal, subcutaneous) for the human sarcoma cell line HS-1 were established in nonobese diabetic/severe-combined immunodeficient mice. Mice transplanted with rAAV-2-EF1alpha-TK/eGFP-transduced and ganciclovir-exposed tumor cells survived > 5 months while in the nontransduced group all mice had died approximately 1 month after inoculation. These data hold promise for further development of rAAV-2-based suicide gene therapy of sarcomas.

Animals↗

[Surgical procedures for bone neoplasms in children].

The treatment of 40 patients with bone tumors have been presented. The primary tumors were located in the following sites: femur (14), tibia (8), fibula (4), humerus (4), scapula (1), clavicle (2), pelvis (5), hand (1). Investigated group were: osteosarcoma (18), Ewing's sarcoma (14), chondrosarcoma (2), fibrosarcoma (1), synovial sarcoma (1), chondroblastoma (4). In the most frequent malignant bone tumors, osteosarcoma and Ewing's sarcoma, unified management was adapted. The treatment was initiated with multidrug chemotherapy and followed by surgery or radiotherapy (Ewing's sarcoma) of the primary site. Surgery was performed in 30 cases: 19 mutilating operations because of the broad local invasion, 11 conservative surgical procedures (limb -- salvage operations). Satisfactory oncological and functional effect can be achieved after limb-salvage surgical procedures in the cases of localized, especially semimalignant bone tumors.

Antineoplastic Combined Chemotherapy Protocols↗

Carcinogenicity of nickel compounds in animals.

A total of 18 nickel compounds were tested for carcinogenicity in male Fischer rats by a single i.m. injection at equivalent dosages (14 mg Ni/rat). Within two years, the following incidences of sarcomas occurred at the injection site: nickel subsulfide (alpha Ni3S2), 100%, crystalline nickel monosulfide (beta NiS), 100%; nickel ferrosulfide (Ni4FeS4), 100%; nickel oxide (NiO), 93%; nickel subselenide (Ni3Se2), 91%; nickel sulfarsenide (NiAsS), 88%; nickel disulfide (NiS2), 86%; nickel subarsenide (Ni5AS2), 85%; nickel dust, 65%; nickel antimonide (NiSb), 59%; nickel telluride (NiTe), 54%; nickel monoselenide (NiSe), 50%; nickel subarsenide (Ni11AS8), 50%; amorphous nickel monosulfide (NiS), 12%; nickel chromate (NiCrO4), 6%; nickel monoarsenide (NiAs), 0%; nickel titanate (NiTiO3), 0%, ferronickel alloy (NiFe1.6), 0%; 84 vehicle controls, 0%. Distant metastases were found in 109 of 180 sarcoma-bearing rats (61%). The nickel-induced sarcomas included rhabdomyosarcomas, 52%, fibrosarcomas, 18%, undifferentiated sarcomas, 13%, osteosarcomas, 8%, and miscellaneous and unclassified sarcomas, 9%. Kendall's rank-correlation test showed that the carcinogenic activities of the compounds were correlated (p = 0.02) with their nickel mass-fractions, but not with dissolution half-times in rat serum or renal cytosol, or with phagocytic indices by rat peritoneal macrophages in vitro. Rank-correlation (p less than 0.0001) was found between the carcinogenic activities and the potencies of the compounds to induce erythrocytosis in rats. The discovery that the carcinogenic activities of particulate nickel compounds are correlated with a physical property, namely the nickel mass-fraction, may help to elucidate the mechanisms of nickel carcinogenesis; the observation that nickel stimulation of erythropoiesis is correlated with carcinogenic activity provides a new in vivo screening test for use in determining the carcinogenic risk of nickel compounds.

Animals↗

Purification and characterization of human lung fibroblast motility-stimulating factor for human soft tissue sarcoma cells: identification as an NH2-terminal fragment of human fibronectin.

Paracrine motogenic factors, including motility cytokines and extracellular matrix molecules secreted by normal cells, can stimulate metastatic cell invasion. Both intact extracellular matrix molecules and their degradative products may exhibit these activities. We have found that human lung fibroblasts produce paracrine motility-stimulating factors for recently established human sarcoma cell strains. We purified the major fibroblast motility-stimulating factor (FMSF) from human lung fibroblast-conditioned medium by sequential heparin affinity chromatography and DEAE anion exchange chromatography. Lysylendopeptidase C digestion of FMSF and sequencing of peptides purified by reverse-phase high-pressure liquid chromatography identified FMSF as an NH2-terminal fragment of human fibronectin. Using SYN-1 sarcoma cells, FMSF predominantly stimulated chemotaxis and some chemokinesis, and it was chemotactic for a variety of human sarcoma cells, including fibrosarcoma, leiomyosarcoma, liposarcoma, synovial sarcoma, and neurofibrosarcoma cells. The FMSF activity present in human lung fibroblast-conditioned medium was completely eliminated by either neutralization or immunodepletion with a rabbit antihuman-fibronectin antibody, thus further confirming that the NH2-terminal fibronectin fragment was the FMSF responsible for the motility stimulation of human soft tissue sarcoma cells. Because human soft tissue sarcomas have a distinctive hematogenous metastatic pattern (predominantly lung), and lung-derived fibroblasts secrete large amounts of FMSF, FMSF and fibronectin may play a role in stimulating sarcoma invasion into lung tissue.

Amino Acid Sequence↗