Abstract
We present a new human osteosarcoma cell line designated OHS-4. These cells showed a high alkaline phosphatase activity that is not regulated by 1,25 dihydroxyvitamin D3. They exhibited a sensitive adenylate cyclase response to parathyroid hormone but not to prostaglandin E2 or human calcitonin. By Northern blot analysis we could detect type I collagen mRNA but none for type III collagen. The cells were able to produce human osteocalcin at a maximum level of 35 ng per million cells when exposed to 2.4 nM 1,25-dihydroxyvitamin D3 for 96 h. We purified this protein from conditioned media using successive chromatography and assessed its identity by partial amino acid sequencing. When injected into nude mice, the cells retained their osteogenic activity and developed calcified tumors. After Von Kossa staining, we observed nonmineralized osteoid deposits and mineralized deposits with a structure similar to that of trabecular bone by light microscopy. On the basis of its osteoblastic characteristics, this new osteosarcoma cell line may represent the human counterpart of the ROS 17/2 cell line. This cell line represents a valuable model for the isolation and characterization of human bone specific proteins.
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- Aubin J. E., Heersche J. N., Merrilees M. J., Sodek J. Isolation of bone cell clones with differences in growth, hormone responses, and extracellular matrix production. J Cell Biol. 1982 Feb;92(2):452–461. doi: 10.1083/jcb.92.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bellows C. G., Aubin J. E., Heersche J. N., Antosz M. E. Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int. 1986 Mar;38(3):143–154. doi: 10.1007/BF02556874. [DOI] [PubMed] [Google Scholar]
- Beresford J. N., Gallagher J. A., Poser J. W., Russell R. G. Production of osteocalcin by human bone cells in vitro. Effects of 1,25(OH)2D3, 24,25(OH)2D3, parathyroid hormone, and glucocorticoids. Metab Bone Dis Relat Res. 1984;5(5):229–234. doi: 10.1016/0221-8747(84)90064-x. [DOI] [PubMed] [Google Scholar]
- Boland C. J., Fried R. M., Tashjian A. H., Jr Measurement of cytosolic free Ca2+ concentrations in human and rat osteosarcoma cells: actions of bone resorption-stimulating hormones. Endocrinology. 1986 Mar;118(3):980–989. doi: 10.1210/endo-118-3-980. [DOI] [PubMed] [Google Scholar]
- Chen T. L., Cone C. M., Feldman D. Effects of 1 alpha,25-dihydroxyvitamin D3 and glucocorticoids on the growth of rat and mouse osteoblast-like bone cells. Calcif Tissue Int. 1983 Sep;35(6):806–811. doi: 10.1007/BF02405127. [DOI] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Chu M. L., Myers J. C., Bernard M. P., Ding J. F., Ramirez F. Cloning and characterization of five overlapping cDNAs specific for the human pro alpha 1(I) collagen chain. Nucleic Acids Res. 1982 Oct 11;10(19):5925–5934. doi: 10.1093/nar/10.19.5925. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chu M. L., Weil D., de Wet W., Bernard M., Sippola M., Ramirez F. Isolation of cDNA and genomic clones encoding human pro-alpha 1 (III) collagen. Partial characterization of the 3' end region of the gene. J Biol Chem. 1985 Apr 10;260(7):4357–4363. [PubMed] [Google Scholar]
- Dedhar S., Argraves W. S., Suzuki S., Ruoslahti E., Pierschbacher M. D. Human osteosarcoma cells resistant to detachment by an Arg-Gly-Asp-containing peptide overproduce the fibronectin receptor. J Cell Biol. 1987 Sep;105(3):1175–1182. doi: 10.1083/jcb.105.3.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eilon G., Perkins J., Viola M. V. Characteristics of a calcitonin-responsive cell line derived from a human osteosarcoma. Cancer Res. 1983 Aug;43(8):3763–3769. [PubMed] [Google Scholar]
- Fodstad O., Brøgger A., Bruland O., Solheim O. P., Nesland J. M., Pihl A. Characteristics of a cell line established from a patient with multiple osteosarcoma, appearing 13 years after treatment for bilateral retinoblastoma. Int J Cancer. 1986 Jul 15;38(1):33–40. doi: 10.1002/ijc.2910380107. [DOI] [PubMed] [Google Scholar]
- Franceschi R. T., James W. M., Zerlauth G. 1 alpha, 25-dihydroxyvitamin D3 specific regulation of growth, morphology, and fibronectin in a human osteosarcoma cell line. J Cell Physiol. 1985 Jun;123(3):401–409. doi: 10.1002/jcp.1041230316. [DOI] [PubMed] [Google Scholar]
- Franceschi R. T., Romano P. R., Park K. Y. Regulation of type I collagen synthesis by 1,25-dihydroxyvitamin D3 in human osteosarcoma cells. J Biol Chem. 1988 Dec 15;263(35):18938–18945. [PubMed] [Google Scholar]
- Fraser J. D., Otawara Y., Price P. A. 1,25-Dihydroxyvitamin D3 stimulates the synthesis of matrix gamma-carboxyglutamic acid protein by osteosarcoma cells. Mutually exclusive expression of vitamin K-dependent bone proteins by clonal osteoblastic cell lines. J Biol Chem. 1988 Jan 15;263(2):911–916. [PubMed] [Google Scholar]
- Fraser J. D., Price P. A. Induction of matrix Gla protein synthesis during prolonged 1,25-dihydroxyvitamin D3 treatment of osteosarcoma cells. Calcif Tissue Int. 1990 Apr;46(4):270–279. doi: 10.1007/BF02555007. [DOI] [PubMed] [Google Scholar]
- Gutierrez G. E., Mundy G. R., Katz M. S. Alterations in hormone-sensitive adenylate cyclase of cloned rat osteosarcoma cells during long-term culture. J Bone Miner Res. 1986 Aug;1(4):319–326. doi: 10.1002/jbmr.5650010404. [DOI] [PubMed] [Google Scholar]
- Heldin C. H., Westermark B., Wasteson A. Chemical and biological properties of a growth factor from human-cultured osteosarcoma cells: resemblance with platelet-derived growth factor. J Cell Physiol. 1980 Nov;105(2):235–246. doi: 10.1002/jcp.1041050207. [DOI] [PubMed] [Google Scholar]
- Lajeunesse D., Frondoza C., Schoffield B., Sacktor B. Osteocalcin secretion by the human osteosarcoma cell line MG-63. J Bone Miner Res. 1990 Sep;5(9):915–922. doi: 10.1002/jbmr.5650050904. [DOI] [PubMed] [Google Scholar]
- Mulkins M. A., Manolagas S. C., Deftos L. J., Sussman H. H. 1,25-Dihydroxyvitamin D3 increases bone alkaline phosphatase isoenzyme levels in human osteogenic sarcoma cells. J Biol Chem. 1983 May 25;258(10):6219–6225. [PubMed] [Google Scholar]
- Murray E., Provvedini D., Curran D., Catherwood B., Sussman H., Manolagas S. Characterization of a human osteoblastic osteosarcoma cell line (SAOS-2) with high bone alkaline phosphatase activity. J Bone Miner Res. 1987 Jun;2(3):231–238. doi: 10.1002/jbmr.5650020310. [DOI] [PubMed] [Google Scholar]
- Pan L. C., Williamson M. K., Price P. A. Sequence of the precursor to rat bone gamma-carboxyglutamic acid protein that accumulates in warfarin-treated osteosarcoma cells. J Biol Chem. 1985 Nov 5;260(25):13398–13401. [PubMed] [Google Scholar]
- Partridge N. C., Alcorn D., Michelangeli V. P., Ryan G., Martin T. J. Morphological and biochemical characterization of four clonal osteogenic sarcoma cell lines of rat origin. Cancer Res. 1983 Sep;43(9):4308–4314. [PubMed] [Google Scholar]
- Price P. A., Parthemore J. G., Deftos L. J. New biochemical marker for bone metabolism. Measurement by radioimmunoassay of bone GLA protein in the plasma of normal subjects and patients with bone disease. J Clin Invest. 1980 Nov;66(5):878–883. doi: 10.1172/JCI109954. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rodan G. A., Heath J. K., Yoon K., Noda M., Rodan S. B. Diversity of the osteoblastic phenotype. Ciba Found Symp. 1988;136:78–91. doi: 10.1002/9780470513637.ch6. [DOI] [PubMed] [Google Scholar]
- Rodan S. B., Imai Y., Thiede M. A., Wesolowski G., Thompson D., Bar-Shavit Z., Shull S., Mann K., Rodan G. A. Characterization of a human osteosarcoma cell line (Saos-2) with osteoblastic properties. Cancer Res. 1987 Sep 15;47(18):4961–4966. [PubMed] [Google Scholar]
- Salomon Y., Londos C., Rodbell M. A highly sensitive adenylate cyclase assay. Anal Biochem. 1974 Apr;58(2):541–548. doi: 10.1016/0003-2697(74)90222-x. [DOI] [PubMed] [Google Scholar]
- Scott D. M., Kent G. N., Cohn D. V. Collagen synthesis in cultured osteoblast-like cells. Arch Biochem Biophys. 1980 May;201(2):384–391. doi: 10.1016/0003-9861(80)90526-3. [DOI] [PubMed] [Google Scholar]
- Spector T. Refinement of the coomassie blue method of protein quantitation. A simple and linear spectrophotometric assay for less than or equal to 0.5 to 50 microgram of protein. Anal Biochem. 1978 May;86(1):142–146. doi: 10.1016/0003-2697(78)90327-5. [DOI] [PubMed] [Google Scholar]
- Spiess Y. H., Price P. A., Deftos J. L., Manolagas S. C. Phenotype-associated changes in the effects of 1,25-dihydroxyvitamin D3 on alkaline phosphatase and bone GLA-protein of rat osteoblastic cells. Endocrinology. 1986 Apr;118(4):1340–1346. doi: 10.1210/endo-118-4-1340. [DOI] [PubMed] [Google Scholar]
- Thein R., Lotan R. Sensitivity of cultured human osteosarcoma and chondrosarcoma cells to retinoic acid. Cancer Res. 1982 Nov;42(11):4771–4775. [PubMed] [Google Scholar]
- Yamane T. Establishment and characterization of cell lines derived from a human osteosarcoma. Clin Orthop Relat Res. 1985 Oct;(199):261–271. [PubMed] [Google Scholar]