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Expression of Bone Morphogenetic Proteins by Osteoinductive and Non-osteoinductive Human Osteosarcoma Cells

Department of Microbiology, Immunology and Molecular Genetics and the Wilkinson Laboratory for Cancer Research

H.H.T. Hsu

Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, Kansas 66160

D.J. Schneider

Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, Kansas 66160

M.P. Sarras, JR

Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas 66160

K. Masuhara

Osaka University Medical School, 2-2-Yamada-oka, Suita 565, Japan

L.F. Bonewald

University of Texas Health Science Center, San Antonio, Texas 78284

H.C. Anderson

Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, Kansas 66160

Freeze-dried Saos-2, human osteosarcoma cells, and extracts of Saos-2 cells contain all components necessary to induce ectopic new bone and marrow when implanted into athymic Nu/Nu nuce. On the other hand, human osteosarcoma cells of the U-2 OS strain failed to induce bone formation under the same experimental conditions. Our aim was to compare the relative expressions of known osteoinductive factors including bone morphogenetic proteins (BMPs) and transforming growth factor-β (TGF-β) in these two cell lines in an attempt to explain the unique bone-inducing ability of the Saos-2 cells. Saos-2 cells expressed mRNA for BMP-1, -2, -3, -4, -6, and TGF-β1. The non-osteoinductive U-2 OS cells expressed BMP-2, -4, -5, -6, and -7 as well as TGF-β1 mRNA, while levels of BMP-1 and BMP-3 mRNA were either not detectable or detectable at a very low level in U-2 OS cells. The presence of BMP-1 and -4 protein was confirmed in Saos-2 cells by immunofluorescence, and TGFβ protein was demonstrated by bioassay in both cell types. These findings suggest that Saos-2 cells are endowed qualitatively and quantitatively with sufficient amounts of many bone morphogenetic proteins-especially BMP-1, -3, and -4—to confer osteoinductivity upon these cells. However, the absence of osteoinductivity in U-2 OS cells, despite significant mRNA expression levels of several bone morphogenetic proteins, suggests that, even though expression of one or more bone morphogenetic proteins may be present, it may not necessarily be sufficient to confer osteoinductivity upon U-2 OS cells. U-2 OS cells may be non-osteoinductive because (1) they contain inhibitors to the BMPs or secrete inhibitory binding proteins, (2) they do not process BMPs correctly, or (3) the BMPs are inappropriately localized and sequestered within the U-2 OS cells. Saos 2 cells may be osteoinductive because (1) they uniquely express BMP-1, (2) they express an appropriate combination of interactive BMPs at appropriate levels, and/or (3) the Saos-2 cells elaborate as-yet-unidentified osteoinductive factor(s).

Key Words: osteoinduction • bone morphogenetic protein • osteosarcoma cell • endochondral bone formation • ectopic bone • transforming growth factor-beta

REFERENCES

• Allen RW, Trach KA, Hoch JA ( 1987). Identification of the 37-kDa protein displaying a variable interaction with the erythroid cell membrane as glyceraldehyde-3-phosphate dehydrogenase. J Biol Chem 262:649-653.[Abstract/Free Full Text]
• Anderson HC, Sugamoto K., Morris DC, Hsu HHT, Hunt T. ( 1992). Bone inducing agent (BIA) from cultured human Saos-2 osteosarcoma cells. Bone & Min 16:49-62.
• Beck LS, Deguzman L., Lee WP, Ku Y., McFatridge LA, Gillett NA, et al. (1991). TGF-β1 induces bone closure of skull defects. J Bone Min Res 6:1257-1265.[Medline] [Order article via Infotrieve]
• Bentley H., Hamdy FC, Hart KA, Seid JM, Williams JL, Johnstone D., et al. (1992). Expression of bone morphogenetic proteins in human prostatic adenocarcinoma and benign prostatic hyperplasia. Br J Cancer 66:1159-1163.[Medline] [Order article via Infotrieve]
• Bentz H., Thompson AY, Armstrong R., Chang RJ, Piez KA, Rosen DM (1991). Transforming growth factor beta 2 enhances the osteoinductive activity of a bovine bone derived fraction containing bone morphogenetic protein-2 and protein-3. Matrix 11:269-275.[Medline] [Order article via Infotrieve]
• Bonewald L., Wakefield L., Oreffo ROC, Escobedo A., Twardzik DR, Mundy GR (1991). Latent forms of transforming growth factor-β (TGF-β) derived from bone cultures: identification of a naturally occurring 100-kDa complex with similarity to recombinant latent TGF-β. Molec Endocrinol 5:741-751.[Abstract/Free Full Text]
• Bostrom K., Watson KE, Horn S., Wortham C., Herman IM, Demer LL (1993). Bone morphogenetic protein expression in human atherosclerotic lesions. J Clin Invest 91:1800-1809.[Medline] [Order article via Infotrieve]
• Celeste AJ, lannazzi JA, Taylor RC, Hewick RM, Rosen V., Wang EA, et al. (1990). Identification of TGF-beta family members present in bone-inductive protein purified from bovine bone. Proc Natl Acad Sci USA 87:9843-9847.[Abstract/Free Full Text]
• Cunningham NS, Paralkar V., Reddi AH (1992). Osteogenin and recombinant bone morphogenetic protein 2B are chemotactic for human monocytes and stimulate transforming growth factor beta-1 mRNA expression. Proc Natl Acad Sci USA 89:11740-11744.[Abstract/Free Full Text]
• Derynck R., Jarrett JA, Chen EY, Eaton DH, Bell JR, Assoian RK, et al. (1985). Human transforming growth factor-β complementary DNA sequence and the expression in normal and transformed cells. Nature 316:701-705.[CrossRef][Medline] [Order article via Infotrieve]
• Doll BA, Towle HJ, Hollinger JO, Reddi AH, Mellonig JT (1990). The osteogenic potential of two composite graft systems using osteogenin. J Periodontol 61:745-750.[Medline] [Order article via Infotrieve]
• Dumermuth E., Sterchi EE, Jiang WP, Wolz RL, Bond JS, Flannery AV, et al. (1991). The Astacin family of metalloendopeptidases. J Biol Chem 266:21381-21385.[Abstract/Free Full Text]
• Harris SE, Harris MA, Mahy P., Sabatini M., Dunn J., Boyce B., et al. (1991). Expression of BMP 4 and BMP 3-like mRNA by human prostate carcinoma cells (abstract). J Bone Min Res 6:S193.
• Harris SE, Harris MA, Mahy P., Wozney J., Feng JQ, Mundy GR (1994). Expression of bone morphogenetic protein messenger RNAs by normal rat and human prostate and prostate cancer cells. Prostate 24:204-211.[Medline] [Order article via Infotrieve]
• Howard JE, Smith JC (1993). Analysis of gastrulation: different types of gastrulation movement are induced by different mesoderminducing factors in Xenopus laevis. Mech Dev 43:37-48.[CrossRef][Medline] [Order article via Infotrieve]
• Jones CM, Lyons KM, Hogan BIM (1991). Involvement of bone morphogenetic protein-4 (BMP-4) and Vgr-1 in morphogenesis and neurogenesis in the mouse. Develop 111:531-542.[Abstract]
• Kawaguchi H., Kurokawa T., Hoshino Y., Kawahara H., Ogata E., Matsumoto T. (1992). Immunohistochemical demonstration of bone morphogenetic protein-2 and transforming growth factor-β in the ossification of the posterior longitudinal ligament of the cervical spine. Spine 17:S33-S36.[Medline] [Order article via Infotrieve]
• Luyten FP, Cuuningham NS, Vukicevic S., Pralkar V., Ripamonti U., Reddi AH (1992). Advances in osteogenin and related bone morphogenetic proteins in bone induction and repair. Acta Orthop Belg 58(Suppl 1):263-267.
• Lyons K., Graycar JL, Hashmi S., Lindquist PB, Chen EY, Hogan BIM, et al. (1989). Vgr-1, a mammalian gene related to Xenopus Vg-1, is a member of the transforming growth factor β gene superfamily. Proc Natl Acad Sci USA 86:4554-4558.[Abstract/Free Full Text]
• Lyons KM, Pelton RW, Hogan BIM (1990). Organogenesis and pattern formation in the mouse: RNA distribution patterns suggest a role for bone morphogenetic protein-2A (BMP-2A). Develop 109:833-844.[Abstract/Free Full Text]
• Mark DE, Hollinger JD, Hastings C. Jr, Chen G., Marden LJ, Reddi AH (1990). Repair of calvarial non-unions by osteogenin, a bone inductive protein. Plast Reconstr Surg 86:623-630.[Medline] [Order article via Infotrieve]
• Masuhara K., Nakase T., Suzuki S., Takaoka K., Matsui M., Anderson HC (1995). Use of monoclonal antibody to detect bone morphogenetic protein-4 (BMP-4). Bone 16:91-96.
• Nakase T., Nomura S., Yoshikawa H., Hashimoto J., Hirota S., Kitamura Y., et al. (1994). Transient and localized expression of bone morphogenetic protein-4 messenger RNA during fracture healing. J Bone Min Res 9:651-659.[Medline] [Order article via Infotrieve]
• Ogawa Y., Schmidt DK, Nathan RM, Armstrong RM, Miller KL, Sawamura SJ, et al. (1992). Bovine bone activin enhances bone morphogenetic protein-induced ectopic bone formation. J Biol Chem 267:14233-14237.[Abstract/Free Full Text]
• Ozkaynak E., Schnegelsberg PNJ, Oppermann H. (1991). Murine osteogenic protein (OP-1): high levels of mRNA in kidney. Biochem Biophys Res Commun 179:116-123.[CrossRef][Medline] [Order article via Infotrieve]
• Ozkaynak E., Schnegelsberg PNJ, Jin DF, Clifford GM, Warren FD, Drier EA, et al. (1992). Osteogenic protein-2. A new member of the transforming growth factor P superfamily expressed early in embryogenesis. J Biol Chem 267:25220-25227.[Abstract/Free Full Text]
• Perides G., Safran RM, Downing LA, Charness ME (1994). Regulation of neural cell adhesion molecule and L1 by the transforming growth factor-beta superfamily. Selective effects of the bone morphogenetic proteins. J Biol Chem 269:765-770.[Abstract/Free Full Text]
• Rogers MB, Rosen V., Wozney JM, Gudas LJ (1992). Bone morphogenetic proteins -2 and -4 are involved in the retinoic acid-induced differentiation of embryonal carcinoma cells. Molec Biol Cell 3:189-196.[Abstract]
• Rosen V., Thies RS (1992). The BMP proteins in bone formation and repair. Trends Genet 8:97-102.[Medline] [Order article via Infotrieve]
• Sambrook J., Fritsch EF, Maniatis T. (1989). Molecular cloning: A laboratory manual. Vol. 1. 2nd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
• Vukicevik S., Latin V., Chen P., Batorsky R., Reddi AH, Sampath TK (1994). Localization of osteogenic protein-1 (bone morphogenetic protein-7) during human embryonic development: high affinity binding to basement membranes. Biochem Biophys Res Commun 198:693-700.[CrossRef][Medline] [Order article via Infotrieve]
• Wang EA, Rosen V., Cordes P., Hewick RM, Kriz MJ, Luxenberg DP, et al. (1988). Purification and characterization of other distinct bone-inducing factors. Proc Natl Acad Sci USA 85:9484-9488.[Abstract/Free Full Text]
• Wozney JM, Rosen V., Celeste AJ, Mitsock LM, Whitters MJ, Kriz RW, et al. (1988). Novel regulators of bone formation: Molecular clones and activities. Science 242:1528-1534.[Abstract/Free Full Text]
• Wozney JM, Rosen V., Byrne M., Celeste AJ, Moutsatsos I., Wang BA ( 1990). Growth factors influencing bone development. J Cell Sci Suppl 13:149-156.[Medline] [Order article via Infotrieve]
• Yasko AW, Lane JM, Fellinger EJ, Rosen V., Wozney JM, Wang EA ( 1992). The healing of segmental bone defects induced by recombinant human bone morphogenetic protein (rhBMP-2)-a radiographic, histological and biomechanical study in rats. J Bone ft Surg 74(A):659-670.

Journal of Dental Research, Vol. 75, No. 7, 1518-1523 (1996)
DOI: 10.1177/00220345960750071301


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