Advertisement

Human Cell

, Volume 30, Issue 2, pp 140–148 | Cite as

Establishment and characterization of a novel uterine carcinosarcoma cell line, TU-ECS-1, with mutations of TP53 and KRAS

  • Yohei Chiba
  • Seiya Sato
  • Hiroaki ItamochiEmail author
  • Yasuko Suga
  • Tomoyuki Fukagawa
  • Nao Oumi
  • Tetsuro Oishi
  • Tasuku Harada
  • Tamotsu Sugai
  • Toru Sugiyama
Cell Line

Abstract

A new human uterine carcinosarcoma (UCS) cell line, TU-ECS-1, was established and characterized. The morphological appearance of the cultured cells was an insular of epithelial-like cells arranged in the form of a jigsaw puzzle and mesenchymal-like cells with a spindle-shaped or fibroblast-like morphology. A relatively high proliferation rate was observed with a doubling time of 18.2 h. The chromosome number ranged from 44 to 49 and had an extra chromosome 12 (trisomy 12). The respective half-maximal inhibitory concentrations of cisplatin, paclitaxel, and doxorubicin were 2.9 µM, 154 nM, and 219 ng/mL, respectively. Mutational analysis revealed that TU-ECS-1 cells have mutations of TP53 in exons 4, 6, and 8 and of KRAS at codon 12 (G12D) in exon 2, which is a mutation hot spot on this gene. Western blot analysis showed that p53 protein was overexpressed in TU-ECS-1 cells. Immunostaining of the cultured cells and in vivo tumors showed that the TU-ECS-1 cells and xenografts were positive for epithelial marker cytokeratin AE1/3 and mesenchymal marker vimentin. These results suggested that TU-ECS-1 cells might have both epithelial and mesenchymal characteristics. This cell line may be useful to study the carcinogenesis of UCS and contribute to the development of novel treatment strategies.

Keywords

Uterine carcinosarcoma Establishment TP53 KRAS Molecular-targeted therapy 

Notes

Acknowledgements

We thank Mr. Noriyuki Yamada of the Department of Pathology at Iwate Medical University School of Medicine for assisting with immunohistochemical analysis. The present study was supported by the Project of Human Resource Development for Cancer from the Ministry of Education, Culture, Sports, Science and Technology of Japan and Leading Center for the Development and Research of Cancer Medicine from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

References

  1. 1.
    Silverberg SG, Major FJ, Blessing JA, et al. Carcinosarcoma (malignant mixed mesodermal tumor) of the uterus. A Gynecologic Oncology Group pathologic study of 203 cases. Int J Gynecol Pathol. 1990;9:1–19.CrossRefPubMedGoogle Scholar
  2. 2.
    Bansal N, Herzog TJ, Seshan VE, et al. Uterine carcinosarcomas and grade 3 endometrioid cancers: evidence for distinct tumor behavior. Obstet Gynecol. 2008;112(1):64–70.CrossRefPubMedGoogle Scholar
  3. 3.
    Desai NB, Kollmeier MA, Makker V, Levine DA, Abu-Rustum NR, Alektiar KM. Comparison of outcomes in early stage uterine carcinosarcoma and uterine serous carcinoma. Gynecol Oncol. 2014;135(1):49–53.CrossRefPubMedGoogle Scholar
  4. 4.
    Gonzalez Bosquet J, Terstriep SA, Cliby WA, et al. The impact of multi-modal therapy on survival for uterine carcinosarcomas. Gynecol Oncol. 2010;116(3):419–23.CrossRefPubMedGoogle Scholar
  5. 5.
    Sutton G, Brunetto VL, Kilgore L, et al. A phase III trial of ifosfamide with or without cisplatin in carcinosarcoma of the uterus: a Gynecologic Oncology Group Study. Gynecol Oncol. 2000;79(2):147–53.CrossRefPubMedGoogle Scholar
  6. 6.
    Yamada SD, Burger RA, Brewster WR, Anton D, Kohler MF, Monk BJ. Pathologic variables and adjuvant therapy as predictors of recurrence and survival for patients with surgically evaluated carcinosarcoma of the uterus. Cancer. 2000;88(12):2782–6.CrossRefPubMedGoogle Scholar
  7. 7.
    Sutton G, Kauderer J, Carson LF, Lentz SS, Whitney CW, Gallion H. Adjuvant ifosfamide and cisplatin in patients with completely resected stage I or II carcinosarcomas (mixed mesodermal tumors) of the uterus: a Gynecologic Oncology Group study. Gynecol Oncol. 2005;96(3):630–4.CrossRefPubMedGoogle Scholar
  8. 8.
    Homesley HD, Filiaci V, Markman M, et al. Phase III trial of ifosfamide with or without paclitaxel in advanced uterine carcinosarcoma: a Gynecologic Oncology Group Study. J Clin Oncol. 2007;25(5):526–31.CrossRefPubMedGoogle Scholar
  9. 9.
    Gadducci A, Cosio S, Romanini A, Genazzani AR. The management of patients with uterine sarcoma: a debated clinical challenge. Crit Rev Oncol/Hematol. 2008;65(2):129–42.CrossRefGoogle Scholar
  10. 10.
    Powell MA, Filiaci VL, Rose PG, et al. Phase II evaluation of paclitaxel and carboplatin in the treatment of carcinosarcoma of the uterus: a Gynecologic Oncology Group study. J Clin Oncol. 2010;28(16):2727–31.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Liu P, Cheng H, Roberts TM, Zhao JJ. Targeting the phosphoinositide 3-kinase pathway in cancer. Nat Rev Drug Discov. 2009;8(8):627–44.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Engelman JA, Luo J, Cantley LC. The evolution of phosphatidylinositol 3-kinases as regulators of growth and metabolism. Nat Rev Genet. 2006;7(8):606–19.CrossRefPubMedGoogle Scholar
  13. 13.
    Brosh R, Rotter V. When mutants gain new powers: news from the mutant p53 field. Nat Rev Cancer. 2009;9(10):701–13.PubMedGoogle Scholar
  14. 14.
    Wada H, Enomoto T, Fujita M, et al. Molecular evidence that most but not all carcinosarcomas of the uterus are combination tumors. Cancer Res. 1997;57(23):5379–85.PubMedGoogle Scholar
  15. 15.
    Growdon WB, Roussel BN, Scialabba VL, et al. Tissue-specific signatures of activating PIK3CA and RAS mutations in carcinosarcomas of gynecologic origin. Gynecol Oncol. 2011;121(1):212–7.CrossRefPubMedGoogle Scholar
  16. 16.
    Bashir S, Jiang G, Joshi A, et al. Molecular alterations of PIK3CA in uterine carcinosarcoma, clear cell, and serous tumors. Int J Gynecol Cancer. 2014;24(7):1262–7.CrossRefPubMedGoogle Scholar
  17. 17.
    Morice P, Leary A, Creutzberg C, Abu-Rustum N, Darai E. Endometrial cancer. Lancet. 2016;387(10023):1094–108.CrossRefPubMedGoogle Scholar
  18. 18.
    Vivanco I, Sawyers CL. The phosphatidylinositol 3-kinase AKT pathway in human cancer. Nat Rev Cancer. 2002;2(7):489–501.CrossRefPubMedGoogle Scholar
  19. 19.
    Thigpen JT, Blessing JA, Orr JW Jr, DiSaia PJ. Phase II trial of cisplatin in the treatment of patients with advanced or recurrent mixed mesodermal sarcomas of the uterus: a Gynecologic Oncology Group Study. Cancer Treat Rep. 1986;70(2):271–4.PubMedGoogle Scholar
  20. 20.
    Curtin JP, Blessing JA, Soper JT, DeGeest K. Paclitaxel in the treatment of carcinosarcoma of the uterus: a gynecologic oncology group study. Gynecol Oncol. 2001;83(2):268–70.CrossRefPubMedGoogle Scholar
  21. 21.
    Omura GA, Major FJ, Blessing JA, et al. A randomized study of adriamycin with and without dimethyl triazenoimidazole carboxamide in advanced uterine sarcomas. Cancer. 1983;52(4):626–32.CrossRefPubMedGoogle Scholar
  22. 22.
    Lacour RA, Euscher E, Atkinson EN, et al. A phase II trial of paclitaxel and carboplatin in women with advanced or recurrent uterine carcinosarcoma. Int J Gynecol Cancer. 2011;21(3):517–22.CrossRefPubMedGoogle Scholar
  23. 23.
    Cantrell LA, Blank SV, Duska LR. Uterine carcinosarcoma: a review of the literature. Gynecol Oncol. 2015;137(3):581–8.CrossRefPubMedGoogle Scholar
  24. 24.
    Cancer Genome Atlas Research Network, Kandoth C, Schultz N, et al. Integrated genomic characterization of endometrial carcinoma. Nature. 2013;497(7447):67–73.CrossRefGoogle Scholar
  25. 25.
    Kandoth C, McLellan MD, Vandin F, et al. Mutational landscape and significance across 12 major cancer types. Nature. 2013;502(7471):333–9.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Ines C, Donia O, Rahma B, et al. Implication of K-ras and p53 in colorectal cancer carcinogenesis in Tunisian population cohort. Tumour Biol. 2014;35(7):7163–75.CrossRefPubMedGoogle Scholar
  27. 27.
    Mandal R, Becker S, Strebhardt K. Stamping out RAF and MEK1/2 to inhibit the ERK1/2 pathway: an emerging threat to anticancer therapy. Oncogene. 2016;35(20):2547–61.CrossRefPubMedGoogle Scholar
  28. 28.
    Ihle NT, Byers LA, Kim ES, et al. Effect of KRAS oncogene substitutions on protein behavior: implications for signaling and clinical outcome. J Natl Cancer Inst. 2012;104(3):228–39.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Juliusson G, Oscier DG, Fitchett M, et al. Prognostic subgroups in B-cell chronic lymphocytic leukemia defined by specific chromosomal abnormalities. N Engl J Med. 1990;323(11):720–4.CrossRefPubMedGoogle Scholar
  30. 30.
    Puiggros A, Blanco G, Espinet B. Genetic abnormalities in chronic lymphocytic leukemia: where we are and where we go. Biomed Res Int. 2014;2014:435983.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Horgas G, Grubisic G, Spaventi S. Trisomy and tetrasomy of the long arm of chromosome 1 in a direct preparation of human endometrial adenocarcinoma. Cancer Genet Cytogenet. 1988;35(2):269–72.CrossRefPubMedGoogle Scholar
  32. 32.
    Gancberg D, Scourneau M, Verdebout JM, Larsimont D, Verhest A. Detection of extra chromosomes 12 by fluorescent in situ hybridization (FISH) in ovarian stromal tumors. Study of 12 cases and review of the literature. Ann Pathol. 2001;21(5):393–8.PubMedGoogle Scholar
  33. 33.
    Liang SB, Sonobe H, Taguchi T, et al. Tetrasomy 12 in ovarian tumors of thecoma-fibroma group: a fluorescence in situ hybridization analysis using paraffin sections. Pathol Int. 2001;51(1):37–42.CrossRefPubMedGoogle Scholar
  34. 34.
    Fletcher JA, Gibas Z, Donovan K, et al. Ovarian granulosa-stromal cell tumors are characterized by trisomy 12. Am J Pathol. 1991;138(3):515–20.PubMedPubMedCentralGoogle Scholar
  35. 35.
    Falisi E, Novella E, Visco C, et al. B-cell receptor configuration and mutational analysis of patients with chronic lymphocytic leukaemia and trisomy 12 reveal recurrent molecular abnormalities. Hematol Oncol. 2014;32(1):22–30.CrossRefPubMedGoogle Scholar

Copyright information

© Japan Human Cell Society and Springer Japan 2016

Authors and Affiliations

  • Yohei Chiba
    • 1
  • Seiya Sato
    • 1
  • Hiroaki Itamochi
    • 1
    Email author
  • Yasuko Suga
    • 1
  • Tomoyuki Fukagawa
    • 1
    • 2
  • Nao Oumi
    • 3
  • Tetsuro Oishi
    • 3
  • Tasuku Harada
    • 3
  • Tamotsu Sugai
    • 2
  • Toru Sugiyama
    • 1
  1. 1.Department of Obstetrics and GynecologyIwate Medical University School of MedicineMoriokaJapan
  2. 2.Department of PathologyIwate Medical University School of MedicineMoriokaJapan
  3. 3.Department of Obstetrics and GynecologyTottori University School of MedicineYonagoJapan

Personalised recommendations