Advertisement

Epigenetic priming sensitizes gastric cancer cells to irinotecan and cisplatin by restoring multiple pathways

  • Hiroshi Moro
  • Naoko Hattori
  • Yoshiaki Nakamura
  • Kana Kimura
  • Toshio Imai
  • Masahiro Maeda
  • Masakazu Yashiro
  • Toshikazu UshijimaEmail author
Original Article
  • 230 Downloads

Abstract

Background

Gastric cancer is heavily influenced by aberrant DNA methylation that alters multiple cancer-related pathways, and may respond to DNA demethylating agents, such as 5-aza-2′-deoxycytidine (5-aza-dC). Here, we aimed to analyze whether 5-aza-dC can sensitize gastric cancer cells to clinically used cytotoxic drugs.

Methods

Ten gastric cancer cell lines were treated with 5-aza-dC for 72 h and their growth was analyzed by conducting WST assay. In vivo effect of the drugs was analyzed using xenografts of OCUM-2 M/SN38 cells. Genome-wide expression and DNA methylation analyses were conducted using microarrays, and biological functions were identified through ingenuity pathway analysis.

Results

The cell lines most resistant to SN38 (an active metabolite of irinotecan), CDDP, PTX, and 5-FU, were identified. 5-Aza-dC pre-treatment of the resistant cell lines decreased the IC50 values for SN38 (TMK1, 226.4 nM to 32.91 nM; 44As3, 128.2 nM to 19.32 nM; OCUM2 M/SN38, 74.43 nM to 16.47 nM) and CDDP (TMK1, 5.05 µM to 2.26 µM; OCUM2 M, 10.79 µM to 2.77 µM), but not PTX and 5-FU. The reactivation of apoptosis-related genes, such as RUNX3, PYCARD, TNF, FAS, and FASLG, was induced by pre-treatment with 5-aza-dC, and the DNA demethylation of promoter CpG islands of RUNX3 and PYCARD was confirmed. In a xenograft model with OCUM2 M/SN38, treatment with 5-aza-dC before irinotecan showed markedly enhanced tumor suppression.

Conclusion

Epigenetic priming with 5-aza-dC can improve the sensitivity of gastric cancer cells to SN38 and CDDP.

Keywords

DNA methylation Resistance Decitabine Irinotecan Cisplatin 

Notes

Funding

This study was supported by AMED under grant number JP19ck0106421.

Compliance with ethical standards

Conflicts of interest

T. U. has received a research grant from Ohara Pharmaceutical Co, Ltd. The other authors declare that they have no conflict of interest.

Ethical approval

All institutional and national guidelines for the care and use of laboratory animals were followed.

Supplementary material

10120_2019_1010_MOESM1_ESM.pdf (105 kb)
Supplementary material 1 (PDF 105 kb)

References

  1. 1.
    Jones PA, Issa JP, Baylin S. Targeting the cancer epigenome for therapy. Nat Rev Genet. 2016;17:630–41.CrossRefGoogle Scholar
  2. 2.
    Ushijima T, Sasako M. Focus on gastric cancer. Cancer Cell. 2004;5:121–5.CrossRefGoogle Scholar
  3. 3.
    Yamashita S, Kishino T, Takahashi T, Shimazu T, Charvat H, Kakugawa Y, et al. Genetic and epigenetic alterations in normal tissues have differential impacts on cancer risk among tissues. Proc Natl Acad Sci USA. 2018;115:1328–33.CrossRefGoogle Scholar
  4. 4.
    Toyota M, Ahuja N, Suzuki H, Itoh F, Ohe-Toyota M, Imai K, et al. Aberrant methylation in gastric cancer associated with the CpG island methylator phenotype. Cancer Res. 1999;59:5438–42.PubMedGoogle Scholar
  5. 5.
    Asada K, Nakajima T, Shimazu T, Yamamichi N, Maekita T, Yokoi C, et al. Demonstration of the usefulness of epigenetic cancer risk prediction by a multicentre prospective cohort study. Gut. 2015;64:388–96.CrossRefGoogle Scholar
  6. 6.
    Maeda M, Nakajima T, Oda I, Shimazu T, Yamamichi N, Maekita T, et al. High impact of methylation accumulation on metachronous gastric cancer: 5-year follow-up of a multicentre prospective cohort study. Gut. 2017;66:1721–3.CrossRefGoogle Scholar
  7. 7.
    Yoda Y, Takeshima H, Niwa T, Kim JG, Ando T, Kushima R, et al. Integrated analysis of cancer-related pathways affected by genetic and epigenetic alterations in gastric cancer. Gastric Cancer. 2015;18:65–76.CrossRefGoogle Scholar
  8. 8.
    Kantarjian H, Issa JP, Rosenfeld CS, Bennett JM, Albitar M, DiPersio J, et al. Decitabine improves patient outcomes in myelodysplastic syndromes: results of a phase III randomized study. Cancer. 2006;106:1794–803.CrossRefGoogle Scholar
  9. 9.
    Diesch J, Zwick A, Garz AK, Palau A, Buschbeck M, Gotze KS. A clinical-molecular update on azanucleoside-based therapy for the treatment of hematologic cancers. Clin Epigenetics. 2016;8:71.CrossRefGoogle Scholar
  10. 10.
    Stresemann C, Lyko F. Modes of action of the DNA methyltransferase inhibitors azacytidine and decitabine. Int J Cancer. 2008;123:8–13.CrossRefGoogle Scholar
  11. 11.
    Juergens RA, Wrangle J, Vendetti FP, Murphy SC, Zhao M, Coleman B, et al. Combination epigenetic therapy has efficacy in patients with refractory advanced non-small cell lung cancer. Cancer Discov. 2011;1:598–607.CrossRefGoogle Scholar
  12. 12.
    Matei D, Fang F, Shen C, Schilder J, Arnold A, Zeng Y, et al. Epigenetic resensitization to platinum in ovarian cancer. Cancer Res. 2012;72:2197–205.CrossRefGoogle Scholar
  13. 13.
    Scandura JM, Roboz GJ, Moh M, Morawa E, Brenet F, Bose JR, et al. Phase 1 study of epigenetic priming with decitabine prior to standard induction chemotherapy for patients with AML. Blood. 2011;118:1472–80.CrossRefGoogle Scholar
  14. 14.
    Halpern AB, Othus M, Huebner EM, Buckley SA, Pogosova-Agadjanyan EL, Orlowski KF, et al. Mitoxantrone, etoposide and cytarabine following epigenetic priming with decitabine in adults with relapsed/refractory acute myeloid leukemia or other high-grade myeloid neoplasms: a phase 1/2 study. Leukemia. 2017;31:2560–7.CrossRefGoogle Scholar
  15. 15.
    Bang YJ, Van Cutsem E, Feyereislova A, Chung HC, Shen L, Sawaki A, et al. Trastuzumab in combination with chemotherapy versus chemotherapy alone for treatment of HER2-positive advanced gastric or gastro-oesophageal junction cancer (ToGA): a phase 3, open-label, randomised controlled trial. Lancet. 2010;376:687–97.CrossRefGoogle Scholar
  16. 16.
    Wilke H, Muro K, Van Cutsem E, Oh SC, Bodoky G, Shimada Y, et al. Ramucirumab plus paclitaxel versus placebo plus paclitaxel in patients with previously treated advanced gastric or gastro-oesophageal junction adenocarcinoma (RAINBOW): a double-blind, randomised phase 3 trial. Lancet Oncol. 2014;15:1224–35.CrossRefGoogle Scholar
  17. 17.
    Cancer Genome Atlas Research Network. Comprehensive molecular characterization of gastric adenocarcinoma. Nature. 2014;513:202–9.CrossRefGoogle Scholar
  18. 18.
    Kang YK, Boku N, Satoh T, Ryu MH, Chao Y, Kato K, et al. Nivolumab in patients with advanced gastric or gastro-oesophageal junction cancer refractory to, or intolerant of, at least two previous chemotherapy regimens (ONO-4538-12, ATTRACTION-2): a randomised, double-blind, placebo-controlled, phase 3 trial. Lancet. 2017;390:2461–71.CrossRefGoogle Scholar
  19. 19.
    Shitara K, Ozguroglu M, Bang YJ, Di Bartolomeo M, Mandala M, Ryu MH, et al. Pembrolizumab versus paclitaxel for previously treated, advanced gastric or gastro-oesophageal junction cancer (KEYNOTE-061): a randomised, open-label, controlled, phase 3 trial. Lancet. 2018;392:123–33.CrossRefGoogle Scholar
  20. 20.
    Kim ST, Cristescu R, Bass AJ, Kim KM, Odegaard JI, Kim K, et al. Comprehensive molecular characterization of clinical responses to PD-1 inhibition in metastatic gastric cancer. Nat Med. 2018;24:1449–58.CrossRefGoogle Scholar
  21. 21.
    Schneider BJ, Shah MA, Klute K, Ocean A, Popa E, Altorki N, et al. Phase I study of epigenetic priming with azacitidine prior to standard neoadjuvant chemotherapy for patients with resectable gastric and esophageal adenocarcinoma: evidence of tumor hypomethylation as an indicator of major histopathologic response. Clin Cancer Res. 2017;23:2673–80.CrossRefGoogle Scholar
  22. 22.
    Zhang X, Yashiro M, Qiu H, Nishii T, Matsuzaki T, Hirakawa K. Establishment and characterization of multidrug-resistant gastric cancer cell lines. Anticancer Res. 2010;30:915–21.PubMedGoogle Scholar
  23. 23.
    Shi L, Reid LH, Jones WD, Shippy R, Warrington JA, Baker SC, et al. The MicroArray Quality Control (MAQC) project shows inter- and intraplatform reproducibility of gene expression measurements. Nat Biotechnol. 2006;24:1151–61.CrossRefGoogle Scholar
  24. 24.
    Shigematsu Y, Niwa T, Yamashita S, Taniguchi H, Kushima R, Katai H, et al. Identification of a DNA methylation marker that detects the presence of lymph node metastases of gastric cancers. Oncol Lett. 2012;4:268–74.CrossRefGoogle Scholar
  25. 25.
    Iida N, Okuda Y, Ogasawara O, Yamashita S, Takeshima H, Ushijima T. MACON: a web tool for computing DNA methylation data obtained by the Illumina Infinium Human DNA methylation BeadArray. Epigenomics. 2018;10:249–58.CrossRefGoogle Scholar
  26. 26.
    Kim JG, Takeshima H, Niwa T, Rehnberg E, Shigematsu Y, Yoda Y, et al. Comprehensive DNA methylation and extensive mutation analyses reveal an association between the CpG island methylator phenotype and oncogenic mutations in gastric cancers. Cancer Lett. 2013;330:33–40.CrossRefGoogle Scholar
  27. 27.
    Bibikova M, Barnes B, Tsan C, Ho V, Klotzle B, Le JM, et al. High density DNA methylation array with single CpG site resolution. Genomics. 2011;98:288–95.CrossRefGoogle Scholar
  28. 28.
    Hur K, Niwa T, Toyoda T, Tsukamoto T, Tatematsu M, Yang HK, et al. Insufficient role of cell proliferation in aberrant DNA methylation induction and involvement of specific types of inflammation. Carcinogenesis. 2011;32:35–41.CrossRefGoogle Scholar
  29. 29.
    Tsai HC, Li H, Van Neste L, Cai Y, Robert C, Rassool FV, et al. Transient low doses of DNA-demethylating agents exert durable antitumor effects on hematological and epithelial tumor cells. Cancer Cell. 2012;21:430–46.CrossRefGoogle Scholar
  30. 30.
    Zong L, Hattori N, Yasukawa Y, Kimura K, Mori A, Seto Y, et al. LINC00162 confers sensitivity to 5-Aza-2′-deoxycytidine via modulation of an RNA splicing protein, HNRNPH1. Oncogene. 2019;38:5281–93.CrossRefGoogle Scholar
  31. 31.
    Cui Y, Hausheer F, Beaty R, Zahnow C, Issa JP, Bunz F, et al. A recombinant reporter system for monitoring reactivation of an endogenously DNA hypermethylated gene. Cancer Res. 2014;74:3834–43.CrossRefGoogle Scholar
  32. 32.
    Chuang LS, Ito K, Ito Y. RUNX family: regulation and diversification of roles through interacting proteins. Int J Cancer. 2013;132:1260–71.CrossRefGoogle Scholar
  33. 33.
    Bae SC, Kolinjivadi AM, Ito Y. Functional relationship between p53 and RUNX proteins. J Mol Cell Biol. 2018;11:224–30.CrossRefGoogle Scholar
  34. 34.
    Salminen A, Kauppinen A, Hiltunen M, Kaarniranta K. Epigenetic regulation of ASC/TMS1 expression: potential role in apoptosis and inflammasome function. Cell Mol Life Sci. 2014;71:1855–64.CrossRefGoogle Scholar
  35. 35.
    Fang F, Cardenas H, Huang H, Jiang G, Perkins SM, Zhang C, et al. Genomic and epigenomic signatures in ovarian cancer associated with resensitization to platinum drugs. Cancer Res. 2018;78:631–44.CrossRefGoogle Scholar
  36. 36.
    Sharma A, Vatapalli R, Abdelfatah E, Wyatt McMahon K, Kerner Z, Guzzetta AA, et al. Hypomethylating agents synergize with irinotecan to improve response to chemotherapy in colorectal cancer cells. PLoS One. 2017;12:e0176139.CrossRefGoogle Scholar
  37. 37.
    Annibaldi A, Meier P. Checkpoints in TNF-induced cell death: implications in inflammation and cancer. Trends Mol Med. 2018;24:49–65.CrossRefGoogle Scholar
  38. 38.
    Falvo JV, Tsytsykova AV, Goldfeld AE. Transcriptional control of the TNF gene. Curr Dir Autoimmun. 2010;11:27–60.CrossRefGoogle Scholar

Copyright information

© The International Gastric Cancer Association and The Japanese Gastric Cancer Association 2019

Authors and Affiliations

  1. 1.Division of EpigenomicsNational Cancer Center Research InstituteTokyoJapan
  2. 2.Course of Advanced Clinical Research of CancerJuntendo University Graduate School of MedicineTokyoJapan
  3. 3.Department of Gastroenterology and Gastrointestinal OncologyNational Cancer Center Hospital EastChibaJapan
  4. 4.Central Animal DivisionNational Cancer Center Research InstituteTokyoJapan
  5. 5.Department of Surgical OncologyOsaka City University Graduate School of MedicineOsakaJapan

Personalised recommendations