Advertisement

Journal of Gastroenterology

, Volume 53, Issue 2, pp 197–207 | Cite as

Esophageal cancer stem cells are suppressed by tranilast, a TRPV2 channel inhibitor

  • Atsushi ShiozakiEmail author
  • Michihiro Kudou
  • Daisuke Ichikawa
  • Hitoshi Fujiwara
  • Hiroki Shimizu
  • Takeshi Ishimoto
  • Tomohiro Arita
  • Toshiyuki Kosuga
  • Hirotaka Konishi
  • Shuhei Komatsu
  • Kazuma Okamoto
  • Yoshinori Marunaka
  • Eigo Otsuji
Original Article—Alimentary Tract

Abstract

Background

Recent evidence suggests that the targeting of membrane proteins specifically activated in cancer stem cells (CSCs) is an important strategy for cancer therapy. The objectives of the present study were to investigate the expression and activity of ion-transport-related molecules in the CSCs of esophageal squamous cell carcinoma.

Methods

Cells exhibiting strong aldehyde dehydrogenase 1 family member A1 (ALDH1A1) activity were isolated from TE8 cells by fluorescence-activated cell sorting, and CSCs were then generated with the sphere formation assay. The gene expression profiles of CSCs were examined by microarray analysis.

Results

Among TE8 cells, ALDH1A1 messenger RNA and protein levels were higher in CSCs than in non-CSCs. The CSCs obtained were resistant to cisplatin and had the ability to redifferentiate. The results of the microarray analysis revealed that the expression of 50 genes encoding plasma membrane proteins was altered in CSCs, whereas that of several genes related to ion channels, including transient receptor potential vanilloid 2 (TRPV2), was upregulated. The TRPV2 inhibitor tranilast was more cytotoxic at a lower concentration in CSCs than in non-CSCs, and effectively decreased the number of tumorspheres. Furthermore, tranilast significantly decreased the cell population that strongly expressed ALDH1A1 among TE8 cells.

Conclusions

The results of the present study suggest that TRPV2 is involved in the maintenance of CSCs, and that its specific inhibitor, tranilast, has potential as a targeted therapeutic agent against esophageal squamous cell carcinoma.

Keywords

Esophageal squamous cell carcinoma Cancer stem cell TRPV2 Tranilast 

Notes

Acknowledgements

This work was supported by a Grant-in-Aid for Scientific Research (C) (26461988) and Grants-in-Aid for Young Scientists (B) (15K19903, 15K19904) from the Japan Society for the Promotion of Science.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

535_2017_1338_MOESM1_ESM.doc (56 kb)
Supplementary material 1 (DOC 55 kb)
535_2017_1338_MOESM2_ESM.doc (132 kb)
Supplementary material 2 (DOC 132 kb)
535_2017_1338_MOESM3_ESM.jpg (507 kb)
Supplementary material 3 (JPEG 506 kb)
535_2017_1338_MOESM4_ESM.jpg (1.1 mb)
Supplementary material 4 (JPEG 1107 kb)
535_2017_1338_MOESM5_ESM.jpg (245 kb)
Supplementary material 5 (JPEG 244 kb)
535_2017_1338_MOESM6_ESM.jpg (873 kb)
Supplementary material 6 (JPEG 873 kb)

References

  1. 1.
    Yuequan J, Shifeng C, Bing Z. Prognostic factors and family history for survival of esophageal squamous cell carcinoma patients after surgery. Ann Thorac Surg. 2010;90:908–13.CrossRefPubMedGoogle Scholar
  2. 2.
    Tachimori Y, Ozawa S, Numasaki H, et al. The registration committee for esophageal cancer of the Japan esophageal society. Comprehensive registry of esophageal cancer in Japan, 2009. Esophagus. 2016;13:110–37.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Kamangar F, Dores GM, Anderson WF. Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol. 2006;24:2137–50.CrossRefPubMedGoogle Scholar
  4. 4.
    Clarke MF, Dick JE, Dirks PB, et al. Cancer stem cells—perspectives on current status and future directions: AACR workshop on cancer stem cells. Cancer Res. 2006;66:9339–44.CrossRefPubMedGoogle Scholar
  5. 5.
    Visvader JE, Lindeman GJ. Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer. 2008;8:755–68.CrossRefPubMedGoogle Scholar
  6. 6.
    Reya T, Morrison SJ, Clarke MF, et al. Stem cells, cancer, and cancer stem cells. Nature. 2001;414:105–11.CrossRefPubMedGoogle Scholar
  7. 7.
    Dean M, Fojo T, Bates S. Tumour stem cells and drug resistance. Nat Rev Cancer. 2005;5:275–84.CrossRefPubMedGoogle Scholar
  8. 8.
    Almanaa TN, Geusz ME, Jamasbi RJ. A new method for identifying stem-like cells in esophageal cancer cell lines. J Cancer. 2013;4:536–48.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Zhang G, Ma L, Xie YK, et al. Esophageal cancer tumorspheres involve cancer stem-like populations with elevated aldehyde dehydrogenase enzymatic activity. Mol Med Rep. 2012;6:519–24.CrossRefPubMedGoogle Scholar
  10. 10.
    Yang L, Ren Y, Yu X, et al. ALDH1A1 defines invasive cancer stem-like cells and predicts poor prognosis in patients with esophageal squamous cell carcinoma. Mod Pathol. 2014;27:775–83.CrossRefPubMedGoogle Scholar
  11. 11.
    Shiozaki A, Ichikawa D, Otsuji E, et al. Cellular physiological approach for treatment of gastric cancer. World J Gastroenterol. 2014;20:11560–6.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Shiozaki A, Ichikawa D, Fujiwara H, et al. Progress in cellular physiological researches on esophageal cancer. J Tumor. 2014;2:241–7.Google Scholar
  13. 13.
    Shiozaki A, Nako Y, Ichikawa D, et al. Role of the Na+/K+/2Cl cotransporter NKCC1 in cell cycle progression in human esophageal squamous cell carcinoma. World J Gastroenterol. 2014;20:6844–59.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Shiozaki A, Takemoto K, Ichikawa D, et al. The K-Cl cotransporter KCC3 as an independent prognostic factor in human esophageal squamous cell carcinoma. Biomed Res Int. 2014;2014:936401.Google Scholar
  15. 15.
    Shimizu H, Shiozaki A, Ichikawa D, et al. The expression and role of aquaporin 5 in esophageal squamous cell carcinoma. J Gastroenterol. 2014;49:655–66.CrossRefPubMedGoogle Scholar
  16. 16.
    Ochi F, Shiozaki A, Ichikawa D, et al. Carbonic anhydrase XII as an independent prognostic factor in advanced esophageal squamous cell carcinoma. J Cancer. 2015;6:922–9.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Takada T, Takata K, Ashihara E. Inhibition of monocarboxylate transporter 1 suppresses the proliferation of glioblastoma stem cells. J Physiol Sci. 2016;66(5):387–96.CrossRefPubMedGoogle Scholar
  18. 18.
    Johnson S, Chen H, Lo PK. In vitro tumorsphere formation assays. Bioprotocol. 2013;3(3):e325.Google Scholar
  19. 19.
    Yue D, Zhang Z, Li J, et al. Transforming growth factor-beta1 promotes the migration and invasion of sphere-forming stem-like cell subpopulations in esophageal cancer. Exp Cell Res. 2015;336:141–9.CrossRefPubMedGoogle Scholar
  20. 20.
    Long A, Giroux V, Whelan KA, et al. WNT10A promotes an invasive and self-renewing phenotype in esophageal squamous cell carcinoma. Carcinogenesis. 2015;36:598–606.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Chen Q, Song S, Wei S, et al. ABT-263 induces apoptosis and synergizes with chemotherapy by targeting stemness pathways in esophageal cancer. Oncotarget. 2015;6:25883–96.PubMedPubMedCentralGoogle Scholar
  22. 22.
    Brungs D, Aghmesheh M, Vine KL, et al. Gastric cancer stem cells: evidence, potential markers, and clinical implications. J Gastroenterol. 2016;51:313–26.CrossRefPubMedGoogle Scholar
  23. 23.
    Ishimoto T, Nagano O, Yae T, et al. CD44 variant regulates redox status in cancer cells by stabilizing the xCT subunit of system xc- and thereby promotes tumor growth. Cancer Cell. 2011;19:387–400.CrossRefPubMedGoogle Scholar
  24. 24.
    Nagano O, Okazaki S, Saya H. Redox regulation in stem-like cancer cells by CD44 variant isoforms. Oncogene. 2013;32:5191–8.CrossRefPubMedGoogle Scholar
  25. 25.
    Shitara K, Doi T, Nagano O, et al. Dose-escalation study for the targeting of CD44v+ cancer stem cells by sulfasalazine in patients with advanced gastric cancer (EPOC1205). Gastric Cancer. 2016;20(2):341–9.CrossRefPubMedGoogle Scholar
  26. 26.
    Peralvarez-Marin A, Donate-Macian P, Gaudet R. What do we know about the transient receptor potential vanilloid 2 (TRPV2) ion channel? FEBS J. 2013;280:5471–87.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Zhou K, Zhang SS, Yan Y, et al. Overexpression of transient receptor potential vanilloid 2 is associated with poor prognosis in patients with esophageal squamous cell carcinoma. Med Oncol. 2014;31:17.CrossRefPubMedGoogle Scholar
  28. 28.
    Liu G, Xie C, Sun F, et al. Clinical significance of transient receptor potential vanilloid 2 expression in human hepatocellular carcinoma. Cancer Genet Cytogenet. 2010;197:54–9.CrossRefPubMedGoogle Scholar
  29. 29.
    Yamada T, Ueda T, Shibata Y, et al. TRPV2 activation induces apoptotic cell death in human T24 bladder cancer cells: a potential therapeutic target for bladder cancer. Urology. 2010;76:509.CrossRefPubMedGoogle Scholar
  30. 30.
    Monet M, Lehen’kyi V, Gackiere F, et al. Role of cationic channel TRPV2 in promoting prostate cancer migration and progression to androgen resistance. Cancer Res. 2010;70:1225–35.CrossRefPubMedGoogle Scholar
  31. 31.
    Darakhshan S, Pour AB. Tranilast: a review of its therapeutic applications. Pharmacol Res. 2015;91:15–28.CrossRefPubMedGoogle Scholar
  32. 32.
    Zhang D, Spielmann A, Wang L, et al. Mast-cell degranulation induced by physical stimuli involves the activation of transient–receptor-potential channel TRPV2. Physiol Res. 2012;61:113–24.PubMedGoogle Scholar
  33. 33.
    Santoni G, Farfariello V, Liberati S, et al. The role of transient receptor potential vanilloid type-2 ion channels in innate and adaptive immune responses. Front Immunol. 2013;4:34.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Prud’homme GJ, Glinka Y, Toulina A, et al. Breast cancer stem-like cells are inhibited by a non-toxic aryl hydrocarbon receptor agonist. PLoS One. 2010;5:e13831.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Subramaniam V, Ace O, Prud’homme GJ, et al. Tranilast treatment decreases cell growth, migration and inhibits colony formation of human breast cancer cells. Exp Mol Pathol. 2011;90:116–22.CrossRefPubMedGoogle Scholar
  36. 36.
    Darakhshan S, Bidmeshkipour A, Mansouri K, et al. The effects of tamoxifen in combination with Tranilast on CXCL12–CXCR4 axis and invasion in breast cancer cell lines. Iran J Pharm Res. 2014;13:683–93.PubMedPubMedCentralGoogle Scholar
  37. 37.
    Darakhshan S, Ghanbari A, Gholami Rad F, et al. Tamoxifen and tranilast show a synergistic effect against breast cancer in vitro. Bratisl Lek Listy. 2015;116:69–73.PubMedGoogle Scholar
  38. 38.
    Yashiro M, Murahashi K, Matsuoka T, et al. Tranilast (N-3,4-dimethoxycinamoyl anthranilic acid): a novel inhibitor of invasion-stimulating interaction between gastric cancer cells and orthotopic fibroblasts. Anticancer Res. 2003;23(5A):3899–904.PubMedGoogle Scholar
  39. 39.
    Murahashi K, Yashiro M, Inoue T, et al. Tranilast and cisplatin as an experimental combination therapy for scirrhous gastric cancer. Int J Oncol. 1998;13:1235–40.PubMedGoogle Scholar
  40. 40.
    Noguchi N, Kawashiri S, Tanaka A, et al. Effects of fibroblast growth inhibitor on proliferation and metastasis of oral squamous cell carcinoma. Oral Oncol. 2003;39:240–7.CrossRefPubMedGoogle Scholar

Copyright information

© Japanese Society of Gastroenterology 2017

Authors and Affiliations

  • Atsushi Shiozaki
    • 1
    Email author
  • Michihiro Kudou
    • 1
  • Daisuke Ichikawa
    • 1
  • Hitoshi Fujiwara
    • 1
  • Hiroki Shimizu
    • 1
  • Takeshi Ishimoto
    • 1
  • Tomohiro Arita
    • 1
  • Toshiyuki Kosuga
    • 1
  • Hirotaka Konishi
    • 1
  • Shuhei Komatsu
    • 1
  • Kazuma Okamoto
    • 1
  • Yoshinori Marunaka
    • 2
    • 3
  • Eigo Otsuji
    • 1
  1. 1.Division of Digestive Surgery, Department of SurgeryKyoto Prefectural University of MedicineKyotoJapan
  2. 2.Department of Molecular Cell Physiology and Bio-Ionomics, Graduate School of Medical ScienceKyoto Prefectural University of MedicineKyotoJapan
  3. 3.Japan Institute for Food Education and HealthSt. Agnes’ UniversityKyotoJapan

Personalised recommendations