Establishment of a New Scirrhous Gastric Cancer Cell Line with FGFR2 Overexpression, OCUM-14

  • Tomohisa Okuno
  • Masakazu YashiroEmail author
  • Go Masuda
  • Shingo Togano
  • Kenji Kuroda
  • Yuichiro Miki
  • Kosei Hirakawa
  • Masahiko Ohsawa
  • Hideki Wanibuchi
  • Masaichi Ohira
Translational Research and Biomarkers



The prognosis of scirrhous gastric carcinoma (SGC), which is characterized by rapid infiltration and proliferation of cancer cells accompanied by extensive stromal fibrosis, is extremely poor. In this study, we report the establishment of a unique SGC cell line from a gastric cancer patient in whom an autopsy was performed.


A new SGC cell line, OCUM-14, was established from malignant ascites of a male patient with SGC. A postmortem autopsy was performed on the patient. Characterization of OCUM-14 cells was analyzed by microscopic examination, reverse transcription polymerase chain reaction, fluorescence in situ hybridization analysis, immunohistochemical examination, CCK-8 assay, and in vivo assay.


OCUM-14 cells grew singly or in clusters, and were floating and round-shaped. Most OCUM-14 cells had many microvilli on their surfaces. The doubling time was 43.1 h, and the subcutaneous inoculation of 1.0 × 107 OCUM-14 cells into mice resulted in 50% tumor formation. mRNA expressions of fibroblast growth factor receptor 2 (FGFR2) and human epidermal growth factor receptor 2 (HER2) were observed in OCUM-14 cells. FGFR2, but not HER2, overexpression was found in OCUM-14 cells. The heterogeneous overexpression of FGFR2 was also found in both the primary tumor and metastatic lesions of the peritoneum, lymph node, bone marrow, and lung of the patient. The FGFR2 inhibitors AZD4547 and BGJ398 significantly decreased the growth of OCUM-14 cells, while paclitaxel and 5-fluorouracil significantly decreased the proliferation of OCUM-14 cells, but cisplatin did not.


A new gastric cancer cell line, OCUM-14, was established from SGC and showed FGFR2 overexpression. OCUM-14 might be useful for elucidating the characteristic mechanisms of SGC and clarifying the effect of FGFR2 inhibitors on SGC.



The authors thank Kayo Tsubota (Osaka City University Graduate School of Medicine) and Hideki Nakagawa (Research support platform of Osaka City University Graduate School of Medicine) for technical assistance. This study was partially supported by JSPS KAKENHI (Grant-in-Aid for Scientific Research B; Grant Nos. JP23390329, JP18H02883), and the Strategic Research Fund of Osaka City University.

Author Contributions

Study concept and design: MY; material collection: GM, MO, and MY; data acquisition: YM, TO, KK, GM; analysis and interpretation of data: YM, and MY; drafting of the manuscript: YM and MY; and study supervision: MY, MO, KH, and MO.


There are no financial or other interests with regard to the submitted manuscript that might be construed as a conflict of interest.

Supplementary material

10434_2018_7145_MOESM1_ESM.tiff (53 kb)
Fig. S1 Growth curve of OCUM-14 cells. The doubling time of OCUM-14 cells was 43.1 h (TIFF 54 kb)
10434_2018_7145_MOESM2_ESM.tiff (226 kb)
Fig. S2 STR profile of OCUM-14. OCUM-14 was matched in < 80% of alleles for 16 loci (TIFF 227 kb)
10434_2018_7145_MOESM3_ESM.tiff (55 kb)
Fig. S3 Effect of trastuzumab on the growth of OCUM-14. The proliferation of OCUM-14 cells was not inhibited by trastuzumab (TIFF 55 kb)


  1. 1.
    Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65(2):87–108.Google Scholar
  2. 2.
    Yashiro M, Chung YS, Kubo T, Hato F, Sowa M. Differential responses of scirrhous and well-differentiated gastric cancer cells to orthotopic fibroblasts. Br J Cancer. 1996;74(7):1096–103.Google Scholar
  3. 3.
    Otsuji E, Kuriu Y, Okamoto K, et al. Outcome of surgical treatment for patients with scirrhous carcinoma of the stomach. Am J Surg. 2004;188(3):327–32.Google Scholar
  4. 4.
    Nashimoto A, Akazawa K, Isobe Y, et al. Gastric cancer treated in 2002 in Japan: 2009 annual report of the JGCA nationwide registry. Gastric Cancer. 2013;16(1):1–27.Google Scholar
  5. 5.
    Yashiro M, Matsuoka T, Ohira M. The significance of scirrhous gastric cancer cell lines: the molecular characterization using cell lines and mouse models. Hum Cell. Epub 6 Jun 2018.
  6. 6.
    Grygielewicz P, Dymek B, Bujak A, et al. Epithelial-mesenchymal transition confers resistance to selective FGFR inhibitors in SNU-16 gastric cancer cells. Gastric Cancer. 2016;19(1):53–62.Google Scholar
  7. 7.
    Matsumoto K, Arao T, Hamaguchi T, et al. FGFR2 gene amplification and clinicopathological features in gastric cancer. Br J Cancer. 2012;106(4):727–32.Google Scholar
  8. 8.
    Papadopoulos KP, El-Rayes BF, Tolcher AW, et al. A Phase 1 study of ARQ 087, an oral pan-FGFR inhibitor in patients with advanced solid tumours. Br J Cancer. 2017;117(11):1592–9.Google Scholar
  9. 9.
    Porta C, Giglione P, Liguigli W, Paglino C. Dovitinib (CHIR258, TKI258): structure, development and preclinical and clinical activity. Fut Oncol. 2015;11(1):39–50.Google Scholar
  10. 10.
    Angevin E, Lopez-Martin JA, Lin CC, et al. Phase I study of dovitinib (TKI258), an oral FGFR, VEGFR, and PDGFR inhibitor, in advanced or metastatic renal cell carcinoma. Clin Cancer Res. 2013;19(5):1257–68.Google Scholar
  11. 11.
    Van Cutsem E, Bang YJ, Mansoor W, et al. A randomized, open-label study of the efficacy and safety of AZD4547 monotherapy versus paclitaxel for the treatment of advanced gastric adenocarcinoma with FGFR2 polysomy or gene amplification. Ann Oncol. 2017;28(6):1316–24.Google Scholar
  12. 12.
    Javle M, Lowery M, Shroff RT, et al. Phase II study of BGJ398 in patients with FGFR-altered advanced cholangiocarcinoma. J Clin Oncol. 2018;36(3):276–82.Google Scholar
  13. 13.
    Tabernero J, Bahleda R, Dienstmann R, et al. Phase I dose-escalation study of JNJ-42756493, an oral pan-fibroblast growth factor receptor inhibitor, in patients with advanced solid tumors. J Clin Oncol. 2015;33(30):3401–8.Google Scholar
  14. 14.
    Michael M, Bang YJ, Park YS, et al. A phase 1 study of LY2874455, an oral selective pan-FGFR inhibitor, in patients with advanced cancer. Target Oncol. 2017;12(4):463–74.Google Scholar
  15. 15.
    Brooks AN, Kilgour E, Smith PD. Molecular pathways: fibroblast growth factor signaling: a new therapeutic opportunity in cancer. Clin Cancer Res. 2012;18(7):1855–62.Google Scholar
  16. 16.
    Kunii K, Davis L, Gorenstein J, et al. FGFR2-amplified gastric cancer cell lines require FGFR2 and Erbb3 signaling for growth and survival. Cancer Res. 2008;68(7):2340–8.Google Scholar
  17. 17.
    Nakamura K, Yashiro M, Matsuoka T, et al. A novel molecular targeting compound as K-samII/FGF-R2 phosphorylation inhibitor, Ki23057, for Scirrhous gastric cancer. Gastroenterology. 2006;131(5):1530–41.Google Scholar
  18. 18.
    Hattori Y, Odagiri H, Nakatani H, et al. K-sam, an amplified gene in stomach cancer, is a member of the heparin-binding growth factor receptor genes. Proc. Natl. Acad. Sci. USA. 1990;87(15):5983–7.Google Scholar
  19. 19.
    Jung K, Park MI, Kim SE, Park SJ. Borrmann type 4 advanced gastric cancer: focus on the development of scirrhous gastric cancer. Clin Endosc. 2016;49(4):336–45.Google Scholar
  20. 20.
    Seabright M. A rapid banding technique for human chromosomes. Lancet. 1971;2(7731):971–2.Google Scholar
  21. 21.
    Capes-Davis A, Reid YA, Kline MC, et al. Match criteria for human cell line authentication: where do we draw the line? Int. J. Cancer. 2013;132(11):2510–9.Google Scholar
  22. 22.
    Yashiro M, Shinto O, Nakamura K, et al. Synergistic antitumor effects of FGFR2 inhibitor with 5-fluorouracil on scirrhous gastric carcinoma. Int. J. Cancer. 2010;126(4):1004–16.Google Scholar
  23. 23.
    Xie L, Su X, Zhang L, et al. FGFR2 gene amplification in gastric cancer predicts sensitivity to the selective FGFR inhibitor AZD4547. Clin. Cancer Res. 2013;19(9):2572–83.Google Scholar
  24. 24.
    Saka H, Kitagawa C, Kogure Y, et al. Safety, tolerability and pharmacokinetics of the fibroblast growth factor receptor inhibitor AZD4547 in Japanese patients with advanced solid tumours: a Phase I study. Investig New Drugs. 2017;35(4):451–62.Google Scholar
  25. 25.
    Guagnano V, Furet P, Spanka C, et al. Discovery of 3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-{6-[4-(4-ethyl-piperazin-1-yl)-phenylamino]-pyrimidin-4-yl}-1-methyl-urea (NVP-BGJ398), a potent and selective inhibitor of the fibroblast growth factor receptor family of receptor tyrosine kinase. J. Med. Chem. 2011;54(20):7066–83.Google Scholar
  26. 26.
    MacLeod RA, Dirks WG, Matsuo Y, Kaufmann M, Milch H, Drexler HG. Widespread intraspecies cross-contamination of human tumor cell lines arising at source. Int J Cancer. 1999;83(4):555–63.Google Scholar
  27. 27.
    Nakazawa K, Yashiro M, Hirakawa K. Keratinocyte growth factor produced by gastric fibroblasts specifically stimulates proliferation of cancer cells from scirrhous gastric carcinoma. Cancer Res. 2003;63(24):8848–52.Google Scholar
  28. 28.
    Liu YJ, Shen D, Yin X, et al. HER2, MET and FGFR2 oncogenic driver alterations define distinct molecular segments for targeted therapies in gastric carcinoma. Br J Cancer. 2014;110(5):1169–78.Google Scholar
  29. 29.
    Toyokawa T, Yashiro M, Hirakawa K. Co-expression of keratinocyte growth factor and K-sam is an independent prognostic factor in gastric carcinoma. Oncol Rep. 2009;21(4):875–80.Google Scholar
  30. 30.
    Hattori Y, Itoh H, Uchino S, et al. Immunohistochemical detection of K-sam protein in stomach cancer. Clin Cancer Res. 1996;2(8):1373–81.Google Scholar
  31. 31.
    Yashiro M, Hirakawa K. Cancer-stromal interactions in scirrhous gastric carcinoma. Cancer Microenviron. 2010;3(1):127–35.Google Scholar
  32. 32.
    Katoh M, Nakagama H. FGF receptors: cancer biology and therapeutics. Med Res Rev. 2014;34(2):280–300.Google Scholar
  33. 33.
    Gavine PR, Mooney L, Kilgour E, et al. AZD4547: an orally bioavailable, potent, and selective inhibitor of the fibroblast growth factor receptor tyrosine kinase family. Cancer Res. 2012;72(8):2045–56.Google Scholar
  34. 34.
    Noda S, Yashiro M, Toyokawa T, et al. Borrmann’s macroscopic criteria and p-Smad2 expression are useful predictive prognostic markers for cytology-positive gastric cancer patients without overt peritoneal metastasis. Ann Surg Oncol. 2011;18(13):3718–25.Google Scholar
  35. 35.
    Oono Y, Kuwata T, Takashima K, et al. Clinicopathological features and endoscopic findings of HER2-positive gastric cancer. Surg Endosc. 2018;32(9):3964–71.Google Scholar
  36. 36.
    Fuyuhiro Y, Yashiro M, Noda S, et al. Myofibroblasts are associated with the progression of scirrhous gastric carcinoma. Exp Ther Med. 2010;1(4):547–51.Google Scholar
  37. 37.
    Kasashima H, Yashiro M, Nakamae H, et al. CXCL1-chemokine (C-X-C Motif) receptor 2 signaling stimulates the recruitment of bone marrow-derived mesenchymal cells into diffuse-type gastric cancer stroma. Am J Pathol. 2016;186(11):3028–39.Google Scholar
  38. 38.
    Fuyuhiro Y, Yashiro M, Noda S, et al. Upregulation of cancer-associated myofibroblasts by TGF-beta from scirrhous gastric carcinoma cells. Br J Cancer. 2011;105(7):996–1001.Google Scholar
  39. 39.
    Fuyuhiro Y, Yashiro M, Noda S, et al. Cancer-associated orthotopic myofibroblasts stimulates the motility of gastric carcinoma cells. Cancer Sci. 2012;103(4):797–805.Google Scholar

Copyright information

© Society of Surgical Oncology 2019

Authors and Affiliations

  • Tomohisa Okuno
    • 1
    • 2
    • 3
  • Masakazu Yashiro
    • 1
    • 2
    • 3
    Email author
  • Go Masuda
    • 1
  • Shingo Togano
    • 1
    • 2
    • 3
  • Kenji Kuroda
    • 1
    • 2
    • 3
  • Yuichiro Miki
    • 1
    • 2
    • 3
  • Kosei Hirakawa
    • 1
  • Masahiko Ohsawa
    • 4
  • Hideki Wanibuchi
    • 5
  • Masaichi Ohira
    • 1
  1. 1.Department of Surgical OncologyOsaka City University Graduate School of MedicineOsaka CityJapan
  2. 2.Molecular Oncology and TherapeuticsOsaka City University Graduate School of MedicineOsaka CityJapan
  3. 3.Cancer Center for Translational ResearchOsaka City University Graduate School of MedicineOsaka CityJapan
  4. 4.Department of Diagnostic PathologyOsaka City University Graduate School of MedicineOsaka CityJapan
  5. 5.Molecular PathologyOsaka City University Graduate School of MedicineOsaka CityJapan

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