tretinoin and Chondrosarcoma

The highly aggressive adult sarcomas are characterized by high levels of matrix metalloproteinase (MMP)-2 and -9, which play crucial roles in tumor invasion and metastasis by degradation of the extracellular membrane leading to cancer cell spread to distal organs. We examined the effect of cytokines, mitogens, inducers and inhibitors on MMP-2 and MMP-9 secretion in chondrosarcoma (SW-1353), fibrosarcoma (HT-1080), liposarcoma (SW-872) and synovial sarcoma (SW-982) cell lines. The selected compounds included natural cytokines and growth factors, as well as chemical compounds applied in therapy of sarcoma and natural compounds that have demonstrated anticancer therapeutic potential. MMP-2 and MMP-9 secretions were analyzed by gelatinase zymography following 24-h exposure to the tested agents and quantitated by densitometry. Fibrosarcoma, chondrosarcoma, liposarcoma and synovial sarcoma showed bands corresponding to MMP-2 and MMP-9 with dose-dependent enhancement of MMP-9 with phorbol 12-myristate 13-acetate (PMA) treatment. In chondrosarcoma cells, tumor necrosis factor (TNF)-α had a stimulatory effect on MMP-9 and insignificant effect on MMP-2 and interleukin (IL)-1β stimulated MMP-9 and MMP-2. In fibrosarcoma and liposarcoma cells, TNF-α had a profound stimulatory effect on MMP-9, but no effect on MMP-2 and in synovial sarcoma an inhibitory effect on MMP-2 and no effect on MMP-9. IL-1β had a slight inhibitory effect on fibrosarcoma, liposarcoma and synovial sarcoma MMP-2 and MMP-9 except for MMP-9 in synovial sarcoma which showed slight stimulation. Lipopolysaccharide (LPS) stimulated expression of MMP-2 in fibrosarcoma and chondrosarcoma while inhibited it in liposarcoma. Doxycycline, epigallocatechin gallate and the nutrient mixture inhibited MMP-2 and MMP-9 in all cell lines. Actinomycin-D, cyclohexamide, retinoic acid, and dexamethasone inhibited MMP-2 and -9 in chondrosarcoma and fibrosarcoma cells. Our results show that cytokines, mitogens, inducers and inhibitors have an up or down regulatory effect on MMP-2 and MMP-9 expression in adult sarcoma cell lines, suggesting these agents may be effective strategies to treat these cancers.

Topics: Anti-Bacterial Agents; Anti-Inflammatory Agents; Antineoplastic Agents; Antioxidants; Carcinogens; Catechin; Cell Line, Tumor; Chondrosarcoma; Dactinomycin; Dexamethasone; Doxycycline; Fibrosarcoma; Humans; Interleukin-1beta; Lipopolysaccharides; Liposarcoma; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Sarcoma; Sarcoma, Synovial; Tetradecanoylphorbol Acetate; Tretinoin; Tumor Necrosis Factor-alpha

Topics: Aggrecans; Animals; Cartilage, Articular; Cattle; Cells, Cultured; Chondroitin Sulfate Proteoglycans; Chondrosarcoma; Extracellular Matrix Proteins; Interleukin-1; Lectins, C-Type; Protease Inhibitors; Proteoglycans; Rats; Tretinoin; Tumor Cells, Cultured

Our studies show that the growth of transplantable chondrosarcoma in rats is inhibited significantly by N-4-(hydroxycarbophenyl) retinamide (RII) at a dosage of 50 mg/kg. RII show a weak ability to induce the differentiation of HL-60 cells at a concentration of 1 umol/L. However, most of the HL-60 cells were induced to differentiate along granulocyte lineage after exposure to RII (1 mumol/L) and S86019 (10 micrograms/ml). Flow cytometry studies indicated that the majority of HL-60 cells were arrested in G1 phase by RII plus S86019. Northern blot analysis clearly demonstrated that c-myc expression was inhibited after treating HL-60 cells with RII plus S86019 for 12 hours. Moreover, thymidylate synthetase mRNA transcription was inhibited in those differentiated cells.

Topics: Animals; Antineoplastic Agents; Cell Cycle; Chondrosarcoma; Female; Gene Expression Regulation, Neoplastic; Genes, myc; Humans; Leukemia, Promyelocytic, Acute; Male; Rats; Tretinoin; Tumor Cells, Cultured

The distribution of cell surface negatively-charged macromolecules was determined electron microscopically on untreated and on retinoic acid (RA)-treated cultured human osteosarcoma Hs791 and chondrosarcoma Hs705 cells using cationized ferritin (CF), an electron-dense marker of anionic sites. Labeling on the surface of prefixed cells was continuous and uniform whether they were grown in the absence or presence of RA. In contrast, CF distribution on unfixed cells was markedly affected by RA; CF labeling of untreated cells occurred in patches and clusters whereas the label on RA-treated cells was continuous, as on prefixed cells. CF labeling of unfixed cells decreased considerably after incubation of the cells either with hyaluronidase or neuraminidase. There was also a reduction in patching and clustering. Changes induced by RA in the apparent membrane microviscosity, in neuraminidase-releasable sialic acid, or in transglutaminase activity could not be related to the effect of RA on CF-induced anionic site redistribution since these characteristics were modulated differently in the two cell lines. In contrast, RA increased the sialylation of specific cell surface membrane glycoproteins on both cell types. These results suggest that RA prevents redistribution of cell surface sialoglycoconjugates and glycosaminoglycans by CF. This effect may be the result of increased sialylation of specific surface components and may be related causally to the suppression of the transformed phenotype in the sarcoma cells.

Topics: Acyltransferases; Anions; Cell Line; Cell Membrane; Chondrosarcoma; Ferritins; Glycoproteins; Humans; Hyaluronoglucosaminidase; Iron-Binding Proteins; Membrane Proteins; Neuraminidase; Osteosarcoma; Receptors, Cell Surface; Transglutaminases; Tretinoin

The effect of beta-all-trans-retinoic acid (RA) on the synthesis of cellular, cell surface, and secreted glycoconjugates by human Hs705 chondrosarcoma and Hs791 osteosarcoma cells was investigated in vitro. Untreated and RA-treated cells were labeled either metabolically with radioactive precursors or by oxidation of externally exposed cell membrane glycoprotein(s) (GP) by treatment with NalO4 or neuraminidase and galactose oxidase followed by reduction with NaB[3H]4. The cells were solubilized and analyzed by polyacrylamide gel electrophoresis followed by fluorography. RA enhanced the labeling of sialic acid and galactose residues on the GP of relative molecular weight(s) (Mr) in the range 95,000-300,000 on the surfaces of both cell types. [3H]glycosamine incorporation into GP with Mr of 100,000, 150,000, and 190,000 in both cell lines was also stimulated. In the Hs705 cells there was also an increase in the labeling of a 290,000-Mr GP. In contrast, [3H]glucosamine incorporation into glycoconjugates greater than 400,000 Mr in both the cells and the conditioned medium of Hs705 cells decreased. The latter glycoconjugates were susceptible to hyaluronidase and chondroitinases. [3H]glucosamine incorporation into a secreted 230,000-Mr GP, identified as fibronectin, was also reduced. Analyses of conditioned media of cells labeled with [35S]methionine or [14C]proline demonstrated that RA decreased the secretion of procollagen chains and fibronectin. Immunofluorescence revealed that RA alters the distribution of cell-associated fibronectin. These results demonstrated that RA increases the glycosylation of specific cellular and cell surface GP and decreases the production of secreted GP and glycosaminoglycans by the sarcoma cells.

Topics: Cell Line; Cell Membrane; Chondrosarcoma; Glucosamine; Glycoproteins; Humans; Methionine; Molecular Weight; Osteosarcoma; Proline; Tretinoin

Murine embryonal carcinoma tumors were induced to differentiate in vivo using retinoic acid. Six mice bearing seven tumors survived more than 100 days after treatment. Histological samples of these tumors showed no residual embryonal carcinoma cells, and, for the most part, they were benign cystic teratomas. Three tumors, in addition to the benign tissue, had solid, mitotically active areas. Two of these tumors upon transplantation gave rise to progressively growing, potentially lethal tumors which have proven to be permanently transplantable cell lines. Using techniques of light and electron microscopy, immunohistochemistry, flow microfluorometry, and cytogenetics, we have characterized these lines. One is a chondrosarcoma, and one is a glioma:chondrosarcoma mixture. Both are chromosomally different from the parent embryonal carcinoma stem cell line, but both were clearly derived from it.

Topics: Animals; Cell Differentiation; Cell Line; Chondrosarcoma; DNA Replication; Glioma; Karyotyping; Mice; Teratoma; Tretinoin

Daily oral treatment with retinoic acid (100 mg/kg bodyweight) induced regression of a transplantable rat chondrosarcoma. In a previous biochemical investigation we have shown that the tissue breakdown is preceded by the loss of proteoglycan. The present study describes the histological changes induced by retinoic acid. A decrease in the intensity of metachromatic staining with toluidine blue was noted already after 1 day and the discoloration was almost complete after 4 days correlating with the loss of proteoglycan. Especially in the perichondrium there was a rapid proliferation of fibroblasts and monocytes. Osteoclast-like cells were missing, but tumor nodules were eroded and split up by penetrating perichondrium. After 4 days of treatment larger necrotic areas were found, initially in the center of tumor nodules only. In other areas the majority of tumorous chondroblasts survived. Tumor nodules appeared partly mesenchyma-like with some fibroblast-like cells suggesting a dedifferentiation of chondroblasts by retinoic acid. we believe that tumor regression induced by retinoic acid involved proteoglycan degradation by chondroblasts themselves and chondroclast-like activity of monocytes and fibroblasts.

Topics: Animals; Chondrosarcoma; Female; Fibroblasts; Monocytes; Necrosis; Neoplasm Transplantation; Proteoglycans; Rats; Sarcoma, Experimental; Staining and Labeling; Time Factors; Tretinoin

The ability of retinoic acid (RA) to inhibit the growth of three cell lines (Te85, Hs781, and Hs791) derived from human osteosarcomas and two cell lines (Hs705 and Hs819) derived from human chondrosarcomas was studied in culture. The exposure to 10(-5) M RA resulted, within 4 days, in changes in both cell morphology and cell growth. RA-treated cells appeared flat and spread on the substratum more than untreated cells, their exponential growth rates decreased, and their saturation densities were markedly reduced. All these effects could be reversed by removal of RA from the growth medium. The various cell lines exhibited differential susceptibility to the growth-inhibitory effect of RA. The most sensitive was the Hs705 chondrosarcoma. The proliferation of these cells was inhibited 50% by 10(-9) M RA and was completely blocked by 10(-5) m RA. In contrast, the concentrations of RA required for 50% inhibition of Hs791, Te85, Hs819, and Hs781 were 10(-7), 2 X 10(-7), 2.5 X 10(-7), and 2 X 10(-6) M, respectively. Only the Te85 and the Hs781 osteosarcoma cells and cells derived from a chondrosarcoma biopsy were able to form colonies in a semisolid medium, and this growth was dramatically inhibited by RA. These results demonstrate that RA can suppress in these mesenchymal tumor cells the expression of morphological and growth properties frequently associated with transformed cells.

Topics: Cell Division; Cell Line; Cell Survival; Chondrosarcoma; Drug Evaluation, Preclinical; Humans; Osteosarcoma; Tretinoin

When treated with a retinoic acid analog (Ro 11-1430), the Swarm rat chondrosarcoma regressed (t 1/2 = 11-12 days) with a rapid removal of tumor proteoglycan, histologic evidence of mineralization, and cartilage proteoglycan synthesis was suppressed down to a value of 1% of the control. During the first 3 weeks of treatment, the newly synthesized proteoglycan was similar both in aggregation and size to the proteoglycan present in the control. However, after 5 weeks of treatment synthesis shifted to a small nonaggregating proteoglycan with longer glycosaminoglycan chains now containing dermatan sulfate, possibly representing a switch in proteoglycan synthesized. Heparan sulfate was also detected. Unlabeled proteoglycan released from the tissue during Ro 11-1430 treatment was large (Kav = 0.25 on Sepharose CL-2B) but incapable of aggregation, suggesting the initial proteolytic cleavage was in or near the hyaluronic acid-binding region of the proteoglycan. Degradative enzyme activity varied during the period of treatment. Since other tissues remained histologically normal during the treatment with Ro 11-1430, this drug may have possible therapeutic value.

Topics: Animals; Chondrosarcoma; Glycosaminoglycans; Heparitin Sulfate; Male; Proteoglycans; Rats; Sarcoma, Experimental; Tretinoin

In in vitro cultures of prepared pieces of transplantable chondrosarcoma from F344 rats, retinoic acid enhanced the release of proteoglycan, with a lag between 6 and 14 hours. The retinoic acid-induced release of proteoglycan was inhibited in vivo and in vitro by cycloheximide, an inhibitor of protein synthesis, and in vitro by actinomycin D and cordycepin, inhibitors of RNA synthesis. These results suggested that the retinoic acid-induced release of proteoglycan depends on de novo RNA and protein synthesis. The proteoglycan release induced by retinoic acid in vitro was blocked by EDTA but not by other proteinase inhibitors (pepstatin, phenylmethylsulfonyl fluoride, and Trasylol) and was completely restored by Zn2+. The degradation of proteoglycan induced by retinoic acid may depend on newly synthesized metal-dependent proteinases.

Topics: Animals; Chondrosarcoma; Cycloheximide; Edetic Acid; Female; In Vitro Techniques; Kinetics; Neoplasm Proteins; Proteoglycans; Rats; Rats, Inbred F344; RNA, Neoplasm; Sarcoma, Experimental; Tretinoin

A new rapid assay has been developed for measurement of the binding of [3H]retinoic acid to cellular retinoic acid-binding protein. The assay, which uses activated charcoal for the separation of bound from unbound retinoic acid, was used to determine the concentration required to inhibit the binding of [3H]retinoic acid to cellular retinoic acid-binding protein by 50% for 18 retinoids with free carboxylic acid groups. Partially purified cellular retinoic acid-binding proteins isolated from rat testes and carcinogen-induced rat mammary tumors were used for these determinations. The following parameters were also determined for some or all of the retinoids: hypervitaminosis A doses; activity against carcinogen-induced mouse skin papillomas; inhibition of growth of a rat chondrosarcoma; inhibition of growth of 3T6 cells; and differentiation of the embryonal carcinoma cell line PCC4.azaIR. While all retinoids that are potent in these biological test systems bind tightly to cellular retinoic acid-binding protein, the converse is not true. The lack of a consistent quantitative correlation between 50% inhibitory concentration and biological activity is probably due to insufficient concentrations of the retinoid in the target tissue or celll, which is a consequence of factors such as absorbability, metabolism, tissue distribution, and pharmacokinetics.

Topics: Animals; Binding, Competitive; Chondrosarcoma; Female; In Vitro Techniques; Male; Mammary Neoplasms, Experimental; Mice; Neoplasms, Experimental; Papilloma; Rats; Retinol-Binding Proteins; Skin Neoplasms; Testis; Tretinoin; Vitamin A

Synthetic aromatic analogs of retinoic acid were administered i.p. and p.o. to Fischer F344 rats bearing a transplantable chondrosarcoma. 35CO4 incorporation into glycosaminoglycans were compared for neoplastic and normal cartilage explants after removal from animals given various analogs. There was a direct relationship between [35S]glycosaminoglycan synthesis by chondrosarcoma chondrocytes and inhibition of tumor growth. The degree of inhibition of [35S]glycosaminoglycan synthesis in the neoplastic cartilage was dependent on the dose of the retinoid administered. At 20-mg/kg/day doses of retinoid for 4 weeks, 35SO4 incorporated into glycosaminoglycan by treated tumor explants was reduced as much as 95%. There was no reduction of [35S] glycosaminoglycan produced in normal costal cartilage of the same animals. Retinoid treatment of 20-mg/kg/day doses for 4 weeks resulted in a 75% reduction in glycosaminoglycan per mg of chondrosarcoma; there was no reduction in costal cartilage glycosaminoglycan. Retinoid (10- to 20-mg/kg/day doses) elevated collagen levels per mg of chondrosarcoma but had no effect on costal cartilage collagen. Combined in vitro and in vivo studies showed that retinoid administration modified neoplastic chondrocyte function but had no measurable effect on normal chondrocyte function.

Topics: Animals; Cartilage; Chondrosarcoma; Collagen; Dose-Response Relationship, Drug; Glycosaminoglycans; Rats; Rats, Inbred F344; Sarcoma, Experimental; Sulfates; Tretinoin; Vitamin A

An aromatic analog of retinoic acid, all-trans-9-(4-methoxy-2,3,6-trimethylphenyl)-3,7-dimethyl-2,4,6,8-nonatetraenoic acid (Ro 10-1670), its ethyl esther (Ro 10-9359), and ethyl amide (Ro 11-1430) have been shown to inhibit the growth of a transplantable rat chondrosarcoma. The inhibition observed occurred over a range of tolerated doses. At the higher tolerated dose levels, significant regressions of already established tumors were observed. All three compounds were active when administered ip for 2 or 4 weeks. The ethyl ester, Ro 10-9359, and the etyl amide, Ro 11-1430, were also active when administered for 4 weeks as dietary admixes. In the latter experiments, both compounds were equally effective at tolerated doses, but Ro 11-1430 was less toxic than Ro 10-9359 at higher doses.

Topics: Animals; Body Weight; Chondrosarcoma; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Female; Neoplasms, Experimental; Rats; Time Factors; Tretinoin; Vitamin A