Establishment and characterization of novel patient-derived extraskeletal osteosarcoma cell line NCC-ESOS1-C1


Extraskeletal osteosarcoma (ESOS) is a rare mesenchymal malignancy producing osteoid and bone in soft tissue without skeletal attachment. ESOS exhibits chemoresistance and poor prognosis, and is distinct from osseous osteosarcoma. The biological characteristics of ESOS are not fully understood, and patient-derived cell lines of ESOS are not available from public cell banks. Here, we established a novel cell line of ESOS and characterized its genetic and biological characteristics as well as examined its response to anti-cancer reagents. The cell line was established using tumor tissue from a 58-year-old female patient with ESOS, and named as NCC-ESOS1-C1. Phenotypes relevant to malignancy such as proliferation and invasion were examined in vitro, and genetic features were evaluated using the NCC Oncopanel assay. The response to inhibitors was monitored by screening of an anti-cancer reagent library. The cells constantly proliferated, showing spheroid formation and invasion capabilities. The NCC Oncopanel revealed the presence of actionable mutations in PIK3CA. Library screening revealed the presence of anti-cancer reagents with significant anti-proliferative effects on NCC-ESOS1-C1 at a low concentration. In conclusion, we established and characterized a novel ESOS cell line, NCC-ESOS1-C1. This cell line will be a useful resource for basic research and preclinical studies.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 99

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3


  1. 1.

    Park SG, Song JY, Song IG, Kim MS, Shin BS. Cutaneous extraskeletal osteosarcoma on the scar of a previous bone graft. Ann Dermatol. 2011;23:S160–4.

  2. 2.

    Sordillo PP, Hajdu SI, Magill GB, Golbey RB. Extraosseous osteogenic sarcoma. A review of 48 patients. Cancer. 1983;51:727–34.

  3. 3.

    Allan CJ, Soule EH. Osteogenic sarcoma of the somatic soft tissues. Clinicopathologic study of 26 cases and review of literature. Cancer. 1971;27:1121–33.

  4. 4.

    Rao U, Cheng A, Didolkar MS. Extraosseous osteogenic sarcoma: clinicopathological study of eight cases and review of literature. Cancer. 1978;41:1488–96.

  5. 5.

    Chung EB, Enzinger FM. Extraskeletal osteosarcoma. Cancer. 1987;60:1132–42.

  6. 6.

    Huvos AG. Osteogenic sarcoma of bones and soft tissues in older persons. A clinicopathologic analysis of 117 patients older than 60 years. Cancer. 1986;57:1442–9.

  7. 7.

    Lee JH, Griffiths WJ, Bottomley RH. Extraosseous osteogenic sarcoma following an intramuscular injection. Cancer. 1977;40:3097–101.

  8. 8.

    Lin SY, Chen WM, Wu HH, Chen WY, Chen TH. Extraosseous osteogenic sarcoma. J Chin Med Assoc: JCMA. 2005;68:542–5.

  9. 9.

    Lee WR, Laurie J, Townsend AL. Fine structure of a radiation-induced osteogenic sarcoma. Cancer. 1975;36:1414–25.

  10. 10.

    Logue JP, Cairnduff F. Radiation induced extraskeletal osteosarcoma. Br J Radiol. 1991;64:171–2.

  11. 11.

    Furrukh M, Qureshi A, Mamoon N, Fatima M. Anterior abdominal wall extraosseous osteosarcoma, occurring 40 years after para-aortic irradiation. BMJ Case Rep. 2017;.

  12. 12.

    Fine G, Stout AP. Osteogenic sarcoma of the extraskeletal soft tissues. Cancer. 1956;9:1027–43.

  13. 13.

    Lee MA, Yi J, Chae JM. Cutaneous osteosarcoma arising from a burn scar. Skeletal Radiol. 2017;46:547–51.

  14. 14.

    Drut R, Barletta L. Osteogenic sarcoma arising in an old burn scar. J Cutan Pathol. 1975;2:302–6.

  15. 15.

    Vanhooteghem O, Theate I. A rare extraskeletal osteosarcoma appearing after 55 years on a large stage 3 burn scar. Case Rep Dermatol Med. 2018;2018:5185604.

  16. 16.

    Arndt CA, Crist WM. Common musculoskeletal tumors of childhood and adolescence. N Engl J Med. 1999;341:342–52.

  17. 17.

    Longhi A, Bielack SS, Grimer R, et al. Extraskeletal osteosarcoma: a European Musculoskeletal Oncology Society study on 266 patients. Eur J Cancer. 2017;74:9–16.

  18. 18.

    Sio TT, Vu CC, Sohawon S, et al. Extraskeletal osteosarcoma: an International Rare Cancer Network Study. Am J Clin Oncol. 2016;39:32–6.

  19. 19.

    Jour G, Wang L, Middha S, et al. The molecular landscape of extraskeletal osteosarcoma: a clinicopathological and molecular biomarker study. J Pathol Clin Res. 2016;2:9–20.

  20. 20.

    Pisters PW, Patel SR, Varma DG, et al. Preoperative chemotherapy for stage IIIB extremity soft tissue sarcoma: long-term results from a single institution. J Clin Oncol. 1997;15:3481–7.

  21. 21.

    Coindre JM, Terrier P, Bui NB, et al. Prognostic factors in adult patients with locally controlled soft tissue sarcoma. A study of 546 patients from the French Federation of Cancer Centers Sarcoma Group. J Clin Oncol. 1996;14:869–77.

  22. 22.

    Ahmad SA, Patel SR, Ballo MT, et al. Extraosseous osteosarcoma: response to treatment and long-term outcome. J Clin Oncol. 2002;20:521–7.

  23. 23.

    Aparicio S, Hidalgo M, Kung AL. Examining the utility of patient-derived xenograft mouse models. Nat Rev Cancer. 2015;15:311–6.

  24. 24.

    Ghandi M, Huang FW, Jane-Valbuena J, et al. Next-generation characterization of the Cancer Cell Line Encyclopedia. Nature. 2019;569:503–8.

  25. 25.

    Rossi G, Manfrin A, Lutolf MP. Progress and potential in organoid research. Nat Rev Genet. 2018;19:671–87.

  26. 26.

    Bairoch A. The cellosaurus, a cell-line knowledge resource. J Biomol Tech: JBT. 2018;29:25–38.

  27. 27.

    Sunami K, Ichikawa H, Kubo T, et al. Feasibility and utility of a panel testing for 114 cancer-associated genes in a clinical setting: a hospital-based study. Cancer Sci. 2019;110:1480–90.

  28. 28.

    Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics. 2009;25:1754–60.

  29. 29.

    Kato M, Nakamura H, Nagai M, et al. A computational tool to detect DNA alterations tailored to formalin-fixed paraffin-embedded samples in cancer clinical sequencing. Genome Med. 2018;10:44.

  30. 30.

    Landrum MJ, Lee JM, Benson M, et al. ClinVar: improving access to variant interpretations and supporting evidence. Nucleic Acids Res. 2018;46:D1062–7.

  31. 31.

    Forbes SA, Bindal N, Bamford S, et al. COSMIC: mining complete cancer genomes in the Catalogue of Somatic Mutations in Cancer. Nucleic Acids Res. 2011;39:D945–50.

  32. 32.

    Nath S, Devi GR. Three-dimensional culture systems in cancer research: focus on tumor spheroid model. Pharmacol Ther. 2016;163:94–108.

  33. 33.

    Maruyama N, Miyoshi Y, Taguchi T, Tamaki Y, Monden M, Noguchi S. Clinicopathologic analysis of breast cancers with PIK3CA mutations in Japanese women. Clin Cancer Res. 2007;13:408–14.

  34. 34.

    Kalinsky K, Jacks LM, Heguy A, et al. PIK3CA mutation associates with improved outcome in breast cancer. Clin Cancer Res. 2009;15:5049–59.

  35. 35.

    Li SY, Rong M, Grieu F, Iacopetta B. PIK3CA mutations in breast cancer are associated with poor outcome. Breast Cancer Res Treat. 2006;96:91–5.

  36. 36.

    Lai YL, Mau BL, Cheng WH, Chen HM, Chiu HH, Tzen CY. PIK3CA exon 20 mutation is independently associated with a poor prognosis in breast cancer patients. Ann Surg Oncol. 2008;15:1064–9.

  37. 37.

    Mangone FR, Bobrovnitchaia IG, Salaorni S, Manuli E, Nagai MA. PIK3CA exon 20 mutations are associated with poor prognosis in breast cancer patients. Clinics (Sao Paulo, Braz). 2012;67:1285–90.

  38. 38.

    Saal LH, Holm K, Maurer M, et al. PIK3CA mutations correlate with hormone receptors, node metastasis, and ERBB2, and are mutually exclusive with PTEN loss in human breast carcinoma. Cancer Res. 2005;65:2554–9.

  39. 39.

    Michelucci A, Di Cristofano C, Lami A, et al. PIK3CA in breast carcinoma: a mutational analysis of sporadic and hereditary cases. Diagn Mol Pathol. 2009;18:200–5.

  40. 40.

    Samuels Y, Wang Z, Bardelli A, et al. High frequency of mutations of the PIK3CA gene in human cancers. Science. 2004;304:554.

  41. 41.

    Campbell IG, Russell SE, Choong DY, et al. Mutation of the PIK3CA gene in ovarian and breast cancer. Cancer Res. 2004;64:7678–81.

  42. 42.

    Ikenoue T, Kanai F, Hikiba Y, et al. Functional analysis of PIK3CA gene mutations in human colorectal cancer. Cancer Res. 2005;65:4562–7.

  43. 43.

    Samuels Y, Diaz LA Jr, Schmidt-Kittler O, et al. Mutant PIK3CA promotes cell growth and invasion of human cancer cells. Cancer Cell. 2005;7:561–73.

Download references


We thank Drs. Y. Minami, K. Shimizu, T. Mori, T. Uehara M. Sugawara, Y. Araki, S. Toki, T. Hirose, J. Sugaya, and Ms. R. Nakano, Division of Musculoskeletal Oncology, National Cancer Center Hospital, for sampling tumor tissue specimens from surgically resected materials. We also appreciate the technical assistance by Mrs. A. Sei, Division of Rare Cancer Research. We would like to thank Editage ( for English-language editing and their constructive comments on the manuscript. This research was financially supported by the National Cancer Center Research and Development Fund (Grant no. 29-A-2), and technically assisted by the Fundamental Innovative Oncology Core in the National Cancer Center.

Author information

Correspondence to Tadashi Kondo.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (XLSX 23 kb)

Supplementary material 2 (XLSX 14 kb)

Supplementary material 3 (XLSX 13 kb)

Supplementary material 4 (XLSX 9 kb)

Supplementary material 5 (XLSX 9 kb)

Supplementary material 6 (XLSX 10 kb)

Supplementary material 7 (XLSX 24 kb)

Supplementary material 8: Supplementary Fig. 1 Representative images of the invasion assay in the cell lines NCC-ESOS1-C1 and MG63. AD Representative images of the invasion assay in NCC-ESOS1-C1 A: 1 × 105 cells after 24 h; B: 2 × 105 cells after 24 h; C: 1 × 105 cells after 48 h; and D: 2 × 105 cells after 48 h. EH: Representative images of the invasion assay in MG63. E: 1 × 105 cells after 24 h; F: 2 × 105 cells after 24 h; G: 1 × 105 cells after 48 h; and H: 2 × 105 cells after 48 h. Scale bar indicates 200 µm. (TIFF 1606 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Kito, F., Oyama, R., Noguchi, R. et al. Establishment and characterization of novel patient-derived extraskeletal osteosarcoma cell line NCC-ESOS1-C1. Human Cell 33, 283–290 (2020).

Download citation


  • Extraskeletal osteosarcoma
  • Patient-derived cancer cell line
  • Drug screening
  • Actionable genes
  • NCC Oncopanel