Immunospecificity of non-histone chromosomal proteins in 89Sr-induced osteogenic sarcoma (mouse).

Antibodies against non-histone chromosomal proteins for 89Sr-induced osteogenic sarcoma (mouse) were prepared by immunization of rabbits. The immunoreactivity of this antigen was then compared with those of non-histone chromosomal proteins from Ehrlich ascites tumor, normal mouse liver, and calf thymus by the method of quantitative microcomplement fixation. The non-histone chromosomal proteins of 98Sr-induced osteogenic sarcoma, fractionated by hydroxylapatite chromatography, exhibited significant affinity for the antibodies. Similar proteins from Ehrlich ascites tumor, normal mouse liver, or calf thymus were virtually inactive, indicating the tissue-specificity of 89Sr-induced osteogenic sarcoma proteins.     

Errors and difficulties in the x-ray diagnosis of osteogenic sarcoma of the long tubular bones

In some cases radiodiagnosis of osteogenic sarcoma is associated with some difficulties resulting from a variety of its x-ray picture. The main factor that influences the x-ray manifestations of osteogenic sarcoma, is a neoplastic bone formation which reflects the morphological essence of this tumor. Other signs depend on a tumor site in one or another anatomical part of the tubular bone, on rates and type ot tumor growth. In some cases the combination of these signs ensures reliable diagnosis. However, along side with typical forms of osteogenic sarcoma there are also other ones which do not fit into the classical concept of this tumor type resulting in diagnostic difficulties and errors.     

Immunotherapy of osteogenic sarcoma with transfer factor

Summary Fifteen patients with osteogenic sarcoma were treated with transfer factor derived from leukocytes of their household contacts. Eight of the fifteen patients were tumor-bearing, and transfer factor therapy was correlated with increased cell-mediated immunity in peripheral blood lymphocytes and with lymphocytic infiltrates into the tumors. There was no marked increased in survival time as compared with historical controls, but this therapy did not accelerate the disease, and there were no untoward side effects.Seven of the fifteen patients were disease-free when transfer therapy was initiated shortly after surgical removal of the primary tumor (five patients) or solitary pulmonary metastases (two patients). These patients received transfer factor injections every 2 weeks for 1–2 years. Six of the seven patients are disease-free 62–82 months after surgery. Compared with probabilities of 5-year survival computed from historical controls, this is significant at P<0.008. Abbreviations used in this paper are: CI, cytotoxicity index; GCT, giant cell tumor of bone; HHC, household contacts; HHC_os, household contacts of patients with osteogenic sarcoma; MIC, mononuclear inflammatory cell; TFCI, transfer factor cytotoxicity index; TSTF, tumor-specific transfer factor.     

Human osteogenic sarcoma cells exhibit enhanced protein phosphorylation

Protein phosphorylation was compared in normal human cells and human osteogenic sarcoma cells. The phosphorylation of endogenous cellular protein substrates was measured by two independent methods, incubation of homogenized cells with [γ-³²P]ATP or labeling of intact cells with Na₂H³²PO₄. Phosphorylated proteins were identified by SDS-polyacrylamide gel electrophoresis and autoradiography. The stained protein bands of all four osteosarcoma cell lines were nearly identical to those of the normal cells. However, each of the osteosarcoma cell lines showed autoradiographic evidence of enhanced phosphorylation in many different protein bands which was neither cyclic AMP-dependent nor a function of cellular growth rate or density. When normal and tumor cell homogenates were mixed prior to incubation with [γ-³²P]ATP, the resulting phosphoprotein patterns resembled those obtained with the tumor cells alone. In addition, a surgically derived osteogenic sarcoma was cultured and an established line obtained; another portion of the fresh tumor was immediately homogenized and used in a phosphorylation assay. The same enhanced phosphorylation pattern was obtained with the homogenized fresh tumor as with the cell line established from it. These results suggest thathuman osteogenic sarcoma cells are able to perform a significantly increased amount of phosphorylation of endegenous cellular protein substrates when compared to normal human cells.     

Osteogenic Sarcoma of the Sella After Radiation Treatment of a Pituitary Adenoma

ObjectiveTo report an uncommon case of osteogenic sarcoma of the sella turcica after radiation treatment of a pituitary adenoma.MethodsWe present the clinical history, physical findings, laboratory data, imaging studies, and pathologic findings in a patient found to have osteogenic sarcoma of the sella after radiation therapy for a nonfunctioning pituitary adenoma.ResultsSix years after transsphenoidal resection and postoperative fractionated radiation therapy for a nonfunctioning pituitary adenoma that extended to the cavernous sinus, a 45-year-old man presented with a sinus infection, diplopia, and ophthalmoplegia of the right eye. A computed tomographic scan of the head showed a mass in the sella with involvement of the optic chiasm and right cavernous sinus. Transsphenoidal resection and debulking of the tumor revealed an osteogenic sarcoma. The patient was discharged from the hospital with residual diplopia and ophthalmoplegia. He was treated with levothyroxine, testosterone, and hydrocortisone. Six weeks later, the patient was readmitted after he was found unresponsive, and computed tomographic scans disclosed a massive cerebrovascular accident. He died a few days later.ConclusionOsteogenic sarcoma is a rare, late complication of radiation treatment of pituitary adenoma. Although radiotherapy remains an effective adjunctive treatment in patients with pituitary adenomas, particularly those with residual or recurrent tumor, potential complications must be acknowledged. (Endocr Pract. 2004;10: 335-338)     

Silencing of HMGA1 expression by RNA interference suppresses growth of osteogenic sarcoma

The expression of high mobility group protein A1 (HMGA1) protein has been closely related to various malignant and prognostic degrees of tumor. To investigate the influence of down-regulating HMGA1 on the tumor and the mechanism underlying antitumor of HMGA1, we transfected the HMGA1 shRNA vector into the osteogenic sarcoma MG-63 cell and observed the changes of cell proliferation, invasion abilities, and the tumor growth. HMGA1 gene expression could be efficiently inhibited, and cell proliferation, migration, invasion, and matrix metalloprotease level were also decreased. BALB/C nude mice injected with the MG-63 cells transfected HMGA1 shRNA showed the significant lower tumor weight, tumor volume, and longer tumor-forming time compared with the control group. Our results suggest that knockdown of HMGA1 could inhibit growth and metastasis potentials of MG-63 cells, which may be a therapeutic target protein for osteogenic sarcoma and may be of biological importance.     

Fine needle aspiration cytology of bone tumors

OBJECTIVE: To study the role of fine needle aspiration cytology (FNAC) in the diagnosis of bone tumors and its impact on therapeutic decisions. STUDY DESIGN: A group of 122 cases of bone tumor were evaluated by FNAC. Detailed diagnoses were compared with the available histology. RESULTS: Diagnostic accuracy of FNAC was 90.5% in this study. FNAC could differentiate between various round cell tumors such as Ewing's sarcoma and myeloma, among various giant cell-rich lesions of bone and between the benign and malignant chondroid bone tumors. Some uncommon variants were also correctly diagnosed. In metastatic bone tumors, the source of primary malignancy could not be indicated in the majority (52.9%) because of the poorly differentiated morphology. Osteoid or osteoid-like material was demonstrable in 63.6% cases of osteogenic sarcoma. A case of chondroblastic osteogenic sarcoma that was reported as chondrosarcoma was the only diagnostic error in the study. FNAC obviated the need of open biopsy in 63.8% patients, and therapeutic decisions were made according to the cytologic diagnoses. CONCLUSION: FNAC plays an important role in the early diagnosis of bone tumors by its accuracy, ease of use and rapidity and is helpful in making the therapeutic decisions.     

Osteogenic sarcoma of the mandible: a plea for radical initial surgery

A total of 24 cases of osteogenic sarcoma of the mandible have been reported to the SEER Program of the National Cancer Institute from 1973 to 1983. Out of 14 patients whose disease was staged as regional, only 9 were eligible for 5-year survival figures and only 2 are alive, for a 23 percent 5-year cure. For those patients staged as localized, only four of six eligible for 5-year cure survived, for a 66 percent 5-year survival. Considering all patients reported to SEER who were eligible for 5-year cure, the rate is 40 percent. We believe this reflects the practice of localized resection of these tumors. At Charity Hospital of Louisiana at New Orleans there have been 10 cases of osteogenic sarcoma of the mandible since 1948. There have been no 5-year survivors for those operated on by other services, usually by local resection. On the Tulane Plastic Surgery Service, a total of six cases of osteogenic sarcoma of the mandible were treated by radical surgery with 100 percent survival from 11 to 22 years.     

Identification of human populations with a high incidence of cellular immunity against breast carcinoma

Summary Cell mediated immunity specifically directed against breast carcinoma, but not against osteogenic sarcoma or other carcinomas, was found to occur in the household contacts of patients with active breast carcinoma (primary and metastatic). Such immunity was not found in the normal population, nor in household contacts of patients with osteogenic sarcoma; it was also not found in household contacts of breast carcinoma patients who had been disease free for two or more years. Patients with breast carcinoma had cytotoxicity indexes (CI) comparable to the normal population, regardless of their clinical condition.     

Antitumor properties of vindesine-monoclonal antibody conjugates

Summary The anticancer alkaloid vindesine (VDS) was conjugated to four mouse monoclonal antibodies recognizing human tumor-associated antigens. The antibodies were 96.5 (antimelanoma, IgG2a); 791T/36 (antiosteogenic sarcoma, IgG2b); 11.285.14, and 14.95.55 (anticarcinoembryonic antigen, IgG1 and IgG2a respectively). Conjugates VDS-96.5 and VDS-791T/36 were tested in vitro and shown to be specifically cytotoxic for target cells expressing the appropriate antigen. The in vivo effects of the antibodies and conjugates were tested against human tumor xenografts in athymic or immunodeprived mice using multiple treatments. Conjugate VDS-96.5 retarded the initial growth of a melanoma xenograft, whereas free antibody was without effect. Similarly, VDS-791T/36 but not free antibody retarded the growth of osteogenic sarcoma 791T. The most marked antitumor effects observed were those obtained with VDS conjugates of the anti-CEA antibodies against a colorectal tumor xenograft. Antibody 14.95.55 suppressed tumor growth both alone and as a VDS conjugate, whereas 11.285.14 produced only a slight effect alone but an almost complete and lasting suppression of tumor growth as a VDS conjugate. Free VDS had little effect at nontoxic levels. Acute studies showed that VDS-11.285.14 conjugate was considerably less toxic than free VDS in Balb/c mice.     

Demonstration of virus particles in Moloney murine sarcoma virus-induced periosteal bone in mice

Balb/c mice were inoculated intramuscularly with Moloney murine sarcoma virus in one of the hind legs. This led to the rapid development of a regressive sarcoma and also to the proliferation and osteogenic differentiation of cells in the adjacent periosteum. Examination of the tissues by transmission electron microscopy revealed the presence of type A and C virus particles within the sarcoma cells as well as within the cells of the newly formed bone. Extracellular type C virus particles were formed by budding from the cell surface and by release from disintegrating cells. No virus particles were found in the bone or the surrounding soft tissues of the contralateral, noninfected leg. These observations suggest that viral infection of periosteal cells are at least partly responsible for the osteogenic response associated with the virus-induced sarcoma. Production of growth factors by the sarcoma cells could also contribute to this process.     

A technique for developing established cell lines from human osteosarcomas.

A method is described which has been successfully used to develop two human osteogenic sarcomas into established lines in culture. This method provides a means whereby cells growing from explanted tumor tissue can be immediately cloned and the fibroblastic (nonneoplastic) cells thus selected against. Both lines have been passaged for over 100 population doublings since cloning and have retained the ability to form colonies from single cells plated at low density without the use of feeder layers or conditioned medium. In culture, the osteogenic sarcoma cells are nonfibroblastic, pile up, and appear to retain a morphological similarity to the in vivo tumors from which they were derived. A karyotype of cells derived from one of the tumors containing a marker chromosome is also presented.     

Cyclic AMP-dependent and -independent effects on tissue-type plasminogen activator activity in osteogenic sarcoma cells; evidence from phosphodiesterase inhibition and parathyroid hormone antagonists

The plasminogen activator (PA) in clonal osteogenic sarcoma cells of rat origin (UMR 106-01 and UMR 106-06) and in osteoblast-rich rat calvarial cells has been characterized using specific antibodies to be tissue-type PA (tPA). An Mr value of 75,000 by SDS-polyacrylamide gel electrophoresis and fibrin autoradiography supports this characterization. There was also evidence for an Mr 105,000 component, which could be due to a proteinase-inhibitor complex. The mechanism of regulation of this tPA activity has been studied in the clonal osteogenic sarcoma cells. Parathyroid hormone (PTH) and prostaglandin E2, which increase cyclic AMP production in the sarcoma cells, also increased tPA activity. The sensitivity and magnitude of the tPA response to PTH and prostaglandin E2 were increased by simultaneous treatment with isobutylmethylxanthine (IBMX) at drug concentrations which had little effect themselves on tPA activity. In UMR 106-06 cells, which unlike UMR 106-01 cells show a cyclic AMP response to calcitonin, tPA activity was also increased in response to calcitonin, and the effect was enhanced by IBMX. 1,25-Dihydroxyvitamin D-3 also increased tPA activity in the cells, but this response was not modified by IBMX. Synthetic peptide antagonists of PTH-responsive adenylate cyclase, [34Tyr]-hPTH (3-34) amide and [34Tyr]-hPTH (5-34) amide, inhibited the PTH-induced increase in tPA activity over the same concentration range at which they inhibited cyclic AMP production, but the antagonist peptides had no effect on the tPA responses to prostaglandin E2, calcitonin or 1,25-dihydroxyvitamin D-3. These data indicate that cyclic AMP mediates the actions of PTH, prostaglandin E2 and calcitonin in increasing tPA activity in the clonal osteogenic sarcoma cells. 1,25-Dihydroxyvitamin D-3, on the other hand, increases tPA activity through a mechanism independent of cyclic AMP.     

Free fibula long bone reconstruction in orthopedic oncology: a surgical algorithm for reconstructive options

The fibula free flap became popular in orthopedic oncology for limb-sparing long bone tumor resection. It is particularly suitable for intercalary or resection arthrodesis options. In the present series, a surgical reconstruction algorithm was used, enabling each patient to receive a personalized technique. During the years 1998 to 2002, 30 patients underwent limb-sparing surgery for long bone sarcoma. There were 18 males and 12 females. Their mean age was 23 years (range, 9 to 70 years). The diagnoses were Ewing's sarcoma (11 patients), osteogenic sarcoma (eight patients), chondrosarcoma (five patients), giant cell tumor of bone (three patients), high-grade soft-tissue sarcoma (two patients), and leiomyosarcoma of bone (one patient). The majority of tumors where located in the lower extremity (23 patients), mostly in the femur (15 patients with four tumors in the proximal femoral shaft, five tumors in the distal femoral shaft, five tumors in the whole femoral shaft, and one tumor in the proximal femoral head). In seven patients, the upper extremity was involved; in six patients, the radius was involved; and in one patient, the humerus was involved. The free fibula flap was used in three types of approaches: vascularized fibula as an osseous flap only (18 patients), a combination of a vascularized fibula flap in conjunction with an allograft (Capanna's technique; 10 patients), and a free double-barreled fibula (two patients). All flaps survived. Postoperatively, all patients were monitored clinically, radiologically, and by radioisotope bone scan studies. Callus formation and union were shown 2.6 to 8 months postoperatively. Patients who underwent lower extremity reconstruction were nonweightbearing for 3 to 9 months, with a transition period in which they used a brace and gradually increased weightbearing until full weightbearing was achieved. Eight patients had 11 recipient-site complications. Two patients (6.7 percent) had hematomas, and three patients (10 percent) had infection and dehiscence of the surgical wound with bone exposure in one patient; all complications resolved with conservative treatment only. Failure of the hardware fixation system occurred in two patients, mandating surgical correction. No fibula donor-site complications were recorded. In intercalary resections, the use of the vascularized fibula flap as an isolated osseous flap might be insufficient. Different body sites have different stress loads to carry, depending on the age of the patient and on his individual physical status. To achieve initial strength in the early period, the authors combined the free fibula flap with an allograft (Capanna's method) or augmented it as a double-barreled fibula. They propose a surgical algorithm to assist the surgeon with the preferred method for reconstruction of various long bone defects in different body locations at childhood or adulthood. Long bone reconstruction using a vascularized fibula flap, alone or in combination with an allograft, autogenous bone graft, or double-barreled fibula for limb-sparing surgery, is a safe and reliable method with a predictable bony union, good functional outcome, and a low complication rate.     

Localization of alkaline phosphatase in subcellular structures of human osteogenic sarcomas.

Electron microscopic study of osteogenic sarcomas has revealed association of the product of the reaction for alkaline phosphatase with membranous structures. The structural and function polymorphism of osteogenic sarcoma cells is also shown.     

Treatment results in osteogenic sarcoma in children and adolescents with the use of karminomycin

Carminomycin, a new antibiotic made in the USSR, was used in the treatment of 21 patients aged 5 to 15 years with extended osteogenic sarcoma. As a result of the treatment the number of the patients with lifetime prolonged for 1-2 years increased from 7.6 to 46 per cent. It was shown that the drug might be used for the prophylaxis of the localized forms of the disease in children.     

The presence of ribonucleic acid within the peripheral zones of two types of mammalian cell

An attempt has been made to locate RNA as a structural component of the peripheries of cultured cells derived from a human osteogenic sarcoma, and L1210 mouse leukaemia cells. In the case of cells derived from the osteogenic sarcoma, their detachment from glass was facilitated by incubation with ribonuclease; on removal from glass, they left cellular “footprints” behind, which were visulized in radioautographs of cells previously labeled with tritiated uridine, and removable with ribonuclease. The electrophoretic data show loss of charge by both types of cell following incubation with ribonuclease. These results are interpreted to indicate that RNA is a structural component of the peripheries of these cells. No attempt is made to speculate on the obvious biological improtance of these observations if they are applicable to cells in general.     

Interferon-α conjugation to human osteogenic sarcoma monoclonal antibody 791T/36

Human lymphoblastoid interferon-alpha (IFN-alpha) has been coupled using N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) to a murine monoclonal antibody (791T/36) which reacts with antigens expressed on human osteogenic sarcomas. The purified conjugates retain antibody activity as defined by their capacity to compete with binding of fluorescein isothiocyanate-labelled 791T/36 antibody to 791T cells. IFN-alpha-791T/36 antibody conjugates synthesized with 125I-trace-labelled IFN-alpha and 131I-trace-labelled antibody also bound to 791T cells, but not to bladder carcinoma T24 cells. The conjugates also retain the capacity of free IFN to activate natural killer cells in human peripheral blood lymphocytes and show specific localization in human osteogenic sarcoma xenografts developing in immunodeprived mice. These findings establish that conjugates containing IFN linked to a monoclonal antibody reacting with osteogenic sarcoma-associated antigens have potential for targeted immunotherapy and in related investigations with antibody has been shown by gamma camera imaging of patients following infusion of 131I-labelled antibody to localize in primary osteogenic sarcomas.     

Mass Spectrometric Identification of Ancient Proteins as Potential Molecular Biomarkers for a 2000-Year-Old Osteogenic Sarcoma

Osteosarcoma is the most common primary malignant tumor of bone usually occurring in young adolescent and children. This disease has a poor prognosis, because of the metastases in the period of tumor progression, which are usually developed previous to the clinical diagnosis. In this paper, a 2000-year-old ancient bone remain with osteogenic sarcoma was analyzed searching for tumor biomarkers which are closely related to this disease. After a specific extraction SDS-PAGE gel electrophoresis followed by tryptic digestion was performed. After the digestion the samples were measured using MALDI TOF/TOF MS. Healthy bone samples from same archaeological site were used as control samples. Our results show that in the pathological skeletal remain several well known tumor biomarkers are detected such as annexin A10, BCL-2-like protein, calgizzarin, rho GTPase-activating protein 7, HSP beta-6 protein, transferrin and vimentin compared to the control samples. The identified protein biomarkers can be useful in the discovery of malignant bone lesions such as osteosarcoma in the very early stage of the disease from paleoanthropological remains.     

Recombinant human bone morphogenetic protein-2 promotes Indian hedgehog-mediated osteo-chondrogenic differentiation of a human chondrocytic cell line in vivo and in vitro

We examined osteo-chondrogenic differentiation of a human chondrocytic cell line (USAC) by rhBMP-2 in vivo and in vitro. USAC was established from a transplanted tumor to athymic mouse derived from an osteogenic sarcoma of the mandible. USAC usually shows chondrocytic phenotypes in vivo and in vitro. rhBMP-2 up-regulated not only the mRNA expression of types II and X collagen, but also the mRNA expression of osteocalcin and Cbfa1 in USAC cells in vitro. In vivo experimental cartilaginous tissue formation was prominent in the chamber with rhBMP-2 when compared with the chamber without rhBMP-2. USAC cells implanted with rhBMP-2 often formed osteoid-like tissues surrounded by osteoblastic cells positive for type I collagen. rhBMP up-regulated Ihh, and the expression of Ihh was well correlated with osteo-chondrogenic cell differentiation. These results suggest that rhBMP-2 promotes chondrogenesis and also induces osteogenic differentiation of USAC cells in vivo and in vitro through up-regulation of Ihh.