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An Orthotopic Model of Human Osteosarcoma Growth and Spontaneous Pulmonary Metastasis

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Abstract

Osteosarcoma is the most common primary malignancy of bone and patients often develop pulmonary metastases. In order to investigate the pathogenesis of human osteosarcoma, there is a great need to develop a clinically relevant animal model. Here we report the development of an osteosarcoma animal model using three related human osteosarcoma lines, the parental TE-85 and two derivative lines MNNG/HOS and 143B. In vitro characterization demonstrated that the 143B line had the greatest cell migration and the least cell adhesion activities among the three lines. The 143B line also exhibited the greatest ability for anchorage independent growth. When GFP-tagged osteosarcoma cells were injected into the proximal tibia of athymic mice, we found that 143B cells were highly tumorigenic and metastatic, and MNNG/HOS cells were tumorigenic but significantly less metastatic. TE85 cells were neither tumorigenic nor metastatic. The number of pulmonary metastases was found 50-fold higher in 143B injected animals than that in MNNG/HOS injected mice. No pulmonary metastases were detected in TE85 injected animals for up to 8 weeks. Primary tumors formed by MNNG/HOS and 143B cells could be visualized by whole body fluorescence imaging, while the pulmonary metastases were visualized on the necropsied samples. The GFP tagged 143B cells (and to a lesser extent, MNNG/HOS cells) were readily recovered from lung metastases. This clinically relevant model of human osteosarcoma provides varying degrees of tumor growth at the primary site and metastatic potential. Thus, this orthotopic model should be a valuable tool to investigate factors that promote or inhibit osteosarcoma growth and/or metastasis.

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Abbreviations

EMEM:

minimum essential medium eagle

FCS:

fetal calf serum

GFP:

green fluorescent protein

hpf:

high power field

PBS:

phosphate buffered saline

References

  1. JS Whelan (1997) ArticleTitleOsteosarcoma Eur J Cancer 33 1611–8 Occurrence Handle10.1016/S0959-8049(97)00251-7 Occurrence Handle9389923

    Article  PubMed  Google Scholar 

  2. MD Murphey MR Robbin GA McRae et al. (1997) ArticleTitleThe many faces of osteosarcoma Radiographics 17 1205–31 Occurrence Handle9308111

    PubMed  Google Scholar 

  3. CA Stiller AW Craft I Corazziari (2001) ArticleTitleSurvival of children with bone sarcoma in Europe since 1978: Results from the EUROCARE study Eur J Cancer 37 760–6 Occurrence Handle10.1016/S0959-8049(01)00004-1 Occurrence Handle11311651

    Article  PubMed  Google Scholar 

  4. OP Solheim G Saeter I Elomaa TA Alvegard (1992) ArticleTitleThe treatment of osteosarcoma: Present trends. The Scandinavian Sarcoma Group experience Ann Oncol 3 (Suppl) 2 S7–11

    Google Scholar 

  5. HJ Mankin FJ Hornicek AE Rosenberg et al. (2004) ArticleTitleSurvival data for 648 patients with osteosarcoma treated at one institution Clin Orthop 429 286–91 Occurrence Handle15577500

    PubMed  Google Scholar 

  6. N Jaffe P Pearson AW Yasko et al. (2003) ArticleTitleSingle and multiple metachronous osteosarcoma tumors after therapy Cancer 98 2457–66 Occurrence Handle10.1002/cncr.11800 Occurrence Handle14635081

    Article  PubMed  Google Scholar 

  7. SJ Kim JA Choi SH Lee et al. (2004) ArticleTitleImaging findings of extrapulmonary metastases of osteosarcoma Clin Imag 28 291–300 Occurrence Handle10.1016/S0899-7071(03)00206-7

    Article  Google Scholar 

  8. L Kager A Zoubek U Potschger et al. (2003) ArticleTitlePrimary metastatic osteosarcoma: presentation and outcome of patients treated on neoadjuvant Cooperative Osteosarcoma Study Group protocols J Clin Oncol 21 2011–8 Occurrence Handle10.1200/JCO.2003.08.132 Occurrence Handle12743156

    Article  PubMed  Google Scholar 

  9. MP Link AM Goorin AW Miser et al. (1986) ArticleTitleThe effect of adjuvant chemotherapy on relapse-free survival in patients with osteosarcoma of the extremity N Engl J Med 314 1600–6 Occurrence Handle3520317

    PubMed  Google Scholar 

  10. WG Ward K Mikaelian F Dorey et al. (1994) ArticleTitlePulmonary metastases of stage IIB extremity osteosarcoma and subsequent pulmonary metastases J Clin Oncol 12 1849–58 Occurrence Handle8083708

    PubMed  Google Scholar 

  11. RC Haydon A Deyrup A Ishikawa et al. (2002) ArticleTitleCytoplasmic and/or nuclear accumulation of the beta-catenin protein is a frequent event in human osteosarcoma Int J Cancer 102 338–42 Occurrence Handle10.1002/ijc.10719 Occurrence Handle12402302

    Article  PubMed  Google Scholar 

  12. R Gorlick P Anderson I Andrulis et al. (2003) ArticleTitleBiology of childhood osteogenic sarcoma and potential targets for therapeutic development: meeting summary Clin Cancer Res 9 5442–53 Occurrence Handle14654523

    PubMed  Google Scholar 

  13. AA Sandberg JA Bridge (2003) ArticleTitleUpdates on the cytogenetics and molecular genetics of bone and soft tissue tumors: Osteosarcoma and related tumors Cancer Genet Cytogenet 145 1–30 Occurrence Handle10.1016/S0165-4608(03)00105-5 Occurrence Handle12885459

    Article  PubMed  Google Scholar 

  14. BH Hoang T Kubo JH Healey et al. (2004) ArticleTitleExpression of LDL receptor-related protein 5 (LRP5) as a novel marker for disease progression in high-grade osteosarcoma Int J Cancer 109 106–11 Occurrence Handle10.1002/ijc.11677 Occurrence Handle14735475

    Article  PubMed  Google Scholar 

  15. EE Pakos PA Kyzas JP Ioannidis (2004) ArticleTitlePrognostic significance of TP53 tumor suppressor gene expression and mutations in human osteosarcoma: a meta-analysis Clin Cancer Res 10 6208–14 Occurrence Handle10.1158/1078-0432.CCR-04-0246 Occurrence Handle15448009

    Article  PubMed  Google Scholar 

  16. O Feugeas N Guriec A Babin-Boilletot et al. (1996) ArticleTitleLoss of heterozygosity of the RB gene is a poor prognostic factor in patients with osteosarcoma J Clin Oncol. 14 467–72 Occurrence Handle8636759

    PubMed  Google Scholar 

  17. G Gamberi MS Benassi T Bohling et al. (1998) ArticleTitleC-myc and c-fos in human osteosarcoma: Prognostic value of mRNA and protein expression Oncology 55 556–63 Occurrence Handle10.1159/000011912 Occurrence Handle9778623

    Article  PubMed  Google Scholar 

  18. R Yokoyama R Schneider-Stock K Radig T Wex A Roessner (1998) ArticleTitleClinicopathologic implications of MDM2, p53 and K-ras gene alterations in osteosarcomas: MDM2 amplification and p53 mutations found in progressive tumors Pathol Res Pract 194 615–21 Occurrence Handle9793960

    PubMed  Google Scholar 

  19. C Khanna J Prehn C Yeung et al. (2000) ArticleTitleAn orthotopic model of murine osteosarcoma with clonally related variants differing in pulmonary metastatic potential Clin Exp Metast 18 261–71 Occurrence Handle10.1023/A:1006767007547

    Article  Google Scholar 

  20. C Khanna J Khan P Nguyen et al. (2001) ArticleTitleMetastasis-associated differences in gene expression in a murine model of osteosarcoma Cancer Res 61 3750–9 Occurrence Handle11325848

    PubMed  Google Scholar 

  21. C Khanna X Wan S Bose et al. (2004) ArticleTitleThe membrane-cytoskeleton linker ezrin is necessary for osteosarcoma metastasis Nat Med 10 182–6 Occurrence Handle10.1038/nm982 Occurrence Handle14704791

    Article  PubMed  Google Scholar 

  22. RM McAllister MB Gardner AE Greene et al. (1971) ArticleTitleCultivation in vitro of cells derived from a human osteosarcoma Cancer 27 397–402 Occurrence Handle5100401

    PubMed  Google Scholar 

  23. JS Rhim DL Putman P Arnstein et al. (1977) ArticleTitleCharacterization of human cells transformed in vitro by N-methyl-N′-nitro-N-nitrosoguanidine Int J Cancer 19 505–10 Occurrence Handle265298

    PubMed  Google Scholar 

  24. PJ Hensler LA Annab JC Barrett OM Pereira-Smith (1994) ArticleTitleA gene involved in control of human cellular senescence on human chromosome 1q Mol Cell Biol 14 2291–7 Occurrence Handle8139534

    PubMed  Google Scholar 

  25. MH Oliver NK Harrison JE Bishop PJ Cole GJ Laurent (1989) ArticleTitleA rapid and convenient assay for counting cells cultured in microwell plates: Application for assessment of growth factors J Cell Sci 92 IssueIDPt 3 513–8 Occurrence Handle2592453

    PubMed  Google Scholar 

  26. HH Luu GP Zagaja Z Dubauskas et al. (1998) ArticleTitleIdentification of a novel metastasis-suppressor region on human chromosome 12 Cancer Res 58 3561–5 Occurrence Handle9721861

    PubMed  Google Scholar 

  27. O Berlin D Samid R Donthineni-Rao et al. (1993) ArticleTitleDevelopment of a novel spontaneous metastasis model of human osteosarcoma transplanted orthotopically into bone of athymic mice Cancer Res 53 4890–5 Occurrence Handle8402677

    PubMed  Google Scholar 

  28. EC McGary A Heimberger L Mills et al. (2003) ArticleTitleA fully human antimelanoma cellular adhesion molecule/MUC18 antibody inhibits spontaneous pulmonary metastasis of osteosarcoma cells in vivo Clin Cancer Res 9 6560–6 Occurrence Handle14695161

    PubMed  Google Scholar 

  29. I Kjonniksen M Winderen O Bruland O Fodstad (1994) ArticleTitleValidity and usefulness of human tumor models established by intratibial cell inoculation in nude rats Cancer Res 54 1715–9 Occurrence Handle8137286

    PubMed  Google Scholar 

  30. W Misdorp AA Hart (1979) ArticleTitleSome prognostic and epidemiologic factors in canine osteosarcoma J Natl Cancer Inst 62 537–45 Occurrence Handle283283

    PubMed  Google Scholar 

  31. KA Hahn AM Legendre HM Schuller (1997) ArticleTitleAmputation and dexniguldipine as treatment for canine appendicular osteosarcoma J Cancer Res Clin Oncol 123 34–8 Occurrence Handle10.1007/BF01212612 Occurrence Handle8996538

    Article  PubMed  Google Scholar 

  32. DM Haines OS Bruland G Matte et al. (1992) ArticleTitleImmunoscintigraphic detection of primary and metastatic spontaneous canine osteosarcoma with F(ab’)2 fragments of osteosarcoma-associated monoclonal antibody TP-1 Anticancer Res 12 2151–7 Occurrence Handle1295462

    PubMed  Google Scholar 

  33. T Nakano M Tani Y Ishibashi et al. (2003) ArticleTitleBiological properties and gene expression associated with metastatic potential of human osteosarcoma Clin Exp Metast 20 665–74 Occurrence Handle10.1023/A:1027355610603

    Article  Google Scholar 

  34. T Shirakawa SC Ko TA Gardner et al. (1998) ArticleTitle In vivo suppression of osteosarcoma pulmonary metastasis with intravenous osteocalcin promoter-based toxic gene therapy Cancer Gene Ther 5 274–80 Occurrence Handle9824046

    PubMed  Google Scholar 

  35. EA Lafleur NV Koshkina J Stewart et al. (2004) ArticleTitleIncreased Fas expression reduces the metastatic potential of human osteosarcoma cells Clin Cancer Res 10 8114–9 Occurrence Handle10.1158/1078-0432.CCR-04-0353 Occurrence Handle15585647

    Article  PubMed  Google Scholar 

  36. AP Shah S Parmar R O’Regan (2003) ArticleTitleRight atrial and ventricular thrombus infiltrated with osteoblastic osteosarcoma J Cardiovasc Pharmacol Ther 8 307–11 Occurrence Handle14740080

    PubMed  Google Scholar 

  37. K Kimura T Nakano YB Park et al. (2002) ArticleTitleEstablishment of human osteosarcoma cell lines with high metastatic potential to lungs and their utilities for therapeutic studies on metastatic osteosarcoma Clin Exp Metast 19 477–85 Occurrence Handle10.1023/A:1020395816633

    Article  Google Scholar 

  38. M Serra K Scotlandi MC Manara et al. (1993) ArticleTitleEstablishment and characterization of multidrug-resistant human osteosarcoma cell lines Anticancer Res 13 323–9 Occurrence Handle8100126

    PubMed  Google Scholar 

  39. SF Jia LL Worth ES Kleinerman (1999) ArticleTitleA nude mouse model of human osteosarcoma lung metastases for evaluating new therapeutic strategies Clin Exp Metast 17 501–6 Occurrence Handle10.1023/A:1006623001465

    Article  Google Scholar 

  40. G Poste IJ Fidler (1980) ArticleTitleThe pathogenesis of cancer metastasis Nature 283 139–46 Occurrence Handle10.1038/283139a0 Occurrence Handle6985715

    Article  PubMed  Google Scholar 

  41. BA Yoshida MM Sokoloff DR Welch CW Rinker-Schaeffer (2000) ArticleTitleMetastasis-suppressor genes: A review and perspective on an emerging field J Natl Cancer Inst 92 1717–30 Occurrence Handle10.1093/jnci/92.21.1717 Occurrence Handle11058615

    Article  PubMed  Google Scholar 

  42. DR Welch PS Steeg CW Rinker-Schaeffer (2000) ArticleTitleMolecular biology of breast cancer metastasis. Genetic regulation of human breast carcinoma metastasis Breast Cancer Res 2 408–16 Occurrence Handle10.1186/bcr87 Occurrence Handle11250734

    Article  PubMed  Google Scholar 

  43. M Kirsch G Schackert PM Black (2004) ArticleTitleMetastasis and angiogenesis Cancer Treat Res 117 285–304 Occurrence Handle15015566

    PubMed  Google Scholar 

  44. J Folkman (2004) ArticleTitleEndogenous angiogenesis inhibitors Apmis 112 496–507 Occurrence Handle10.1111/j.1600-0463.2004.apm11207-0809.x Occurrence Handle15563312

    Article  PubMed  Google Scholar 

  45. J Folkman (2002) ArticleTitleRole of angiogenesis in tumor growth and metastasis Semin Oncol 29 15–8 Occurrence Handle10.1053/sonc.2002.37263

    Article  Google Scholar 

  46. F Pompetti P Rizzo RM Simon et al. (1996) ArticleTitleOncogene alterations in primary, recurrent, and metastatic human bone tumors J Cell Biochem 63 37–50 Occurrence Handle10.1002/(SICI)1097-4644(199610)63:1<37::AID-JCB3>3.0.CO;2-0 Occurrence Handle8891902

    Article  PubMed  Google Scholar 

  47. M Conzelmann U Linnemann MR Berger (2004) ArticleTitleK-ras codon 12 and 13 mutations are correlated with differential patterns of tumor cell dissemination in colorectal cancer patients Int J Oncol 24 1537–44 Occurrence Handle15138598

    PubMed  Google Scholar 

  48. T Uemura K Hibi T Kaneko et al. (2004) ArticleTitleDetection of K-ras mutations in the plasma DNA of pancreatic cancer patients J Gastroenterol 39 56–60 Occurrence Handle10.1007/s00535-003-1245-1 Occurrence Handle14767735

    Article  PubMed  Google Scholar 

  49. CP Dieterle M Conzelmann U Linnemann MR Berger (2004) ArticleTitleDetection of isolated tumor cells by polymerase chain reaction-restriction fragment length polymorphism for K-ras mutations in tissue samples of 199 colorectal cancer patients Clin Cancer Res 10 641–50 Occurrence Handle10.1158/1078-0432.CCR-1355-02 Occurrence Handle14760087

    Article  PubMed  Google Scholar 

  50. K Okudela H Hayashi T Ito et al. (2004) ArticleTitleK-ras gene mutation enhances motility of immortalized airway cells and lung adenocarcinoma cells via Akt activation: possible contribution to non-invasive expansion of lung adenocarcinoma Am J Pathol 164 91–100 Occurrence Handle14695323

    PubMed  Google Scholar 

  51. Z Fu PC Smith L Zhang et al. (2003) ArticleTitleEffects of raf kinase inhibitor protein expression on suppression of prostate cancer metastasis J Natl Cancer Inst 95 878–89 Occurrence Handle12813171

    PubMed  Google Scholar 

  52. ET Keller Z Fu K Yeung M Brennan (2004) ArticleTitleRaf kinase inhibitor protein: a prostate cancer metastasis suppressor gene Cancer Lett 207 131–7 Occurrence Handle10.1016/j.canlet.2004.02.006 Occurrence Handle15151133

    Article  PubMed  Google Scholar 

  53. G Odabaei D Chatterjee AR Jazirehi et al. (2004) ArticleTitleRaf-1 kinase inhibitor protein: structure, function, regulation of cell signaling, and pivotal role in apoptosis Adv Cancer Res 91 169–200 Occurrence Handlefull_text||10.1016/S0065-230X(04)91005-6 Occurrence Handle15327891

    Article  PubMed  Google Scholar 

  54. N Trakul MR Rosner (2005) ArticleTitleModulation of the MAP kinase signaling cascade by Raf kinase inhibitory protein Cell Res 15 19–23 Occurrence Handle15686621

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, 5841 South Maryland Avenue, Chicago, IL, 60637, USA

    Hue H. Luu, Quan Kang, Jong Kyung Park, Weike Si, Qing Luo, Wei Jiang, Hong Yin, Anthony G. Montag, Michael A. Simon, Terrance D. Peabody, Rex C. Haydon & Tong-Chuan He

  2. The Children’s Hospital of Chongqing University of Medical Sciences, Chongqing, China

    Quan Kang & Qing Luo

  3. Department of Surgery, St. Paul’s Hospital, College of Medicine, The Catholic University of Korea, Seoul, Korea

    Jong Kyung Park

  4. Department of Clinical Biochemistry, The Third Military Medical University, Chongqing, China

    Weike Si

  5. Department of Pathology, The University of Chicago Hospitals, Chicago, Illinois, USA

    Anthony G. Montag

  6. Urological Research Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, Illinois

    Carrie W. Rinker-Schaeffer

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  1. Hue H. Luu
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  2. Quan Kang
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  3. Jong Kyung Park
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  4. Weike Si
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  13. Tong-Chuan He
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Correspondence to Tong-Chuan He.

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Luu, H.H., Kang, Q., Park, J.K. et al. An Orthotopic Model of Human Osteosarcoma Growth and Spontaneous Pulmonary Metastasis. Clin Exp Metastasis 22, 319–329 (2005). https://doi.org/10.1007/s10585-005-0365-9

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  • DOI: https://doi.org/10.1007/s10585-005-0365-9

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