Skip to main content

Advertisement

SpringerLink
Log in

Discovery of YS-1 as a cell line of gastric inflammatory cancer-associated fibroblasts

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Background

Inflammatory cancer-associated fibroblasts (iCAFs) was first identified by co-culture of pancreatic stellate cells and tumor organoids. The key feature of iCAFs is IL-6high/αSMAlow. We examine this phenomenon in gastric cancer using two cell lines of gastric fibroblasts (HGF and YS-1).

Methods and results

HGF or YS-1 were co-cultured with MKN7 (a gastric adenocarcinoma cell line) in Matrigel. IL-6 protein levels in the culture supernatant were measured by ELISA. The increased production of IL-6 was not observed in any of the combinations. Instead, the supernatant of YS-1 exhibited the higher levels of IL-6. YS-1 showed IL-6high/αSMA (ACTA2)low in real-time PCR, mRNA-seq and immunohistochemistry. In mRNA-seq, iCAFs-associated genes and signaling pathways were up-regulated in YS-1. No transition to myofibroblastic phenotype was observed by monolayer culture, or the exposure to sonic hedgehog (SHH) or TGF-β. YS-1 conditioned medium induced changes of morphology and stem-ness/differentiation in NUGC-3 (a human gastric adenocarcinoma cell line) and UBE6T-15 (a human bone marrow-derived mesenchymal stem cell line).

Conclusions

YS-1 is a stable cell line of gastric iCAFs. This discovery will promote further research on iCAFs for many researchers.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Data availability

All data generated or analyzed during this study are included in this article. The raw data of mRNA-seq will be shared on reasonable request to the corresponding author.

References

  1. Öhlund D, Handly-Santana A, Biffi G et al (2017) Distinct populations of inflammatory fibroblasts and myofibroblasts in pancreatic cancer. J Exp Med 214:579–596

    Article  PubMed  PubMed Central  Google Scholar 

  2. Chen J, Huang X-F (2009) Interleukin-6 promotes carcinogenesis through multiple Signal pathways. Dig Dis Sci 54:1373–1374

    Article  PubMed  Google Scholar 

  3. Elyada E, Bolisetty M, Laise P et al (2019) Cross-species single-cell analysis of pancreatic ductal adenocarcinoma reveals Antigen-Presenting Cancer-Associated fibroblasts. Cancer Discov 9:1102–1123

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Houthuijzen JM, de Bruijn R, van der Burg E et al (2023) CD26-negative and CD26-positive tissue-resident fibroblasts contribute to functionally distinct CAF subpopulations in breast cancer. Nat Commun 14:183

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Chen Z, Zhou L, Liu L et al (2020) Single-cell RNA sequencing highlights the role of inflammatory cancer-associated fibroblasts in bladder urothelial carcinoma. Nat Commun 11:1–12

    Article  Google Scholar 

  6. Du YH, Cao J, Jiang X et al (2021) Comprehensive analysis of CXCL12 expression reveals the significance of inflammatory fibroblasts in bladder cancer carcinogenesis and progression. Cancer Cell Int 21:1–15

    Article  Google Scholar 

  7. Chen H, Yang W, Xue X, Li Y, Jin Z, Ji Z (2022) Integrated Analysis revealed an Inflammatory Cancer-Associated fibroblast-based subtypes with Promising implications in Predicting the prognosis and immunotherapeutic response of bladder Cancer patients. Int J Mol Sci 23:15970

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Wang Z, Zhang H, Zhai Y, Li F, Shi X, Ying M (2022) Single-cell profiling reveals heterogeneity of primary and Lymph Node Metastatic Tumors and Immune Cell populations and discovers important prognostic significance of CCDC43 in oral squamous cell carcinoma. Front Immunol 13:1–13

    Google Scholar 

  9. Yang W, Zhang S, Li T, Zhou Z, Pan J (2022) Single-cell analysis reveals that cancer-associated fibroblasts stimulate oral squamous cell carcinoma invasion via the TGF-beta/Smad pathway. Acta Biochim Biophys Sin (Shanghai) 55:262–273

    Article  PubMed  Google Scholar 

  10. Nicolas AM, Pesic M, Engel E et al (2022) Inflammatory fibroblasts mediate resistance to neoadjuvant therapy in rectal cancer. Cancer Cell 40:168–184e13

    Article  CAS  PubMed  Google Scholar 

  11. Peng Z, Ye M, Ding H, Feng Z, Hu K (2022) Spatial transcriptomics atlas reveals the crosstalk between cancer-associated fibroblasts and tumor microenvironment components in colorectal cancer. J Transl Med 20:1–13

    Article  Google Scholar 

  12. Tran LL, Dang T, Thomas R, Rowley DR (2021) ELF3 mediates IL-1α induced differentiation of mesenchymal stem cells to inflammatory iCAFs. Stem Cells 39:1766–1777

    Article  CAS  PubMed  Google Scholar 

  13. Li X, Sun Z, Peng G et al (2022) Single-cell RNA sequencing reveals a pro-invasive cancer-associated fibroblast subgroup associated with poor clinical outcomes in patients with gastric cancer. Theranostics 12:620–638

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Wu YS, Chung I, Wong WF, Masamune A, Sim MS, Looi CY (2017) Paracrine IL-6 signaling mediates the effects of pancreatic stellate cells on epithelial-mesenchymal transition via Stat3/Nrf2 pathway in pancreatic cancer cells. Biochim Biophys Acta Gen Subj 1861:296–306

    Article  CAS  PubMed  Google Scholar 

  15. Shi Y, Gao W, Lytle NK et al (2019) Targeting LIF-mediated paracrine interaction for pancreatic cancer therapy and monitoring. Nature 569:131–135

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Garg B, Giri B, Modi S et al (2018) NFκB in pancreatic stellate cells reduces infiltration of tumors by cytotoxic T cells and killing of Cancer cells, via Up-regulation of CXCL12. Gastroenterology 155:880–891e8

    Article  CAS  PubMed  Google Scholar 

  17. Hu B, Wu C, Mao H et al (2022) Subpopulations of cancer-associated fibroblasts link the prognosis and metabolic features of pancreatic ductal adenocarcinoma. Ann Transl Med 10:262

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Biffi G, Oni TE, Spielman B et al (2019) IL1-Induced JAK/STAT signaling is antagonized by TGFβ to shape CAF heterogeneity in pancreatic ductal adenocarcinoma. Cancer Discov 9:282–301

    Article  PubMed  Google Scholar 

  19. Steele NG, Biffi G, Kemp SB et al (2021) Inhibition of hedgehog signaling alters fibroblast composition in pancreatic Cancer. Clin Cancer Res 27:2023–2037

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Kanda Y (2013) Investigation of the freely available easy-to-use software EZR for medical statistics. Bone Marrow Transpl 48:452–458

    Article  CAS  Google Scholar 

  21. Geng X, Chen H, Zhao L et al (2021) Cancer-Associated Fibroblast (CAF) heterogeneity and targeting therapy of CAFs in pancreatic Cancer. Front Cell Dev Biol 9:1–14

    Article  Google Scholar 

  22. Miyazaki Y, Oda T, Mori N, Kida YS (2020) Adipose-derived mesenchymal stem cells differentiate into pancreatic cancer-associated fibroblasts in vitro. FEBS Open Bio 10:2268–2281

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Peiffer R, Boumahd Y, Gullo C et al (2023) Cancer-Associated Fibroblast Diversity shapes Tumor Metabolism in Pancreatic Cancer. Cancers (Basel) 15:61

    Article  CAS  Google Scholar 

  24. Sunami Y, Chen Y, Trojanowicz B et al (2022) Single cell analysis of cultivated fibroblasts from chronic pancreatitis and pancreatic Cancer patients. Cells 11:2583

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Seeneevassen L, Martin OCB, Lehours P, Dubus P, Varon C (2022) Leukaemia inhibitory factor in gastric cancer: friend or foe? Gastric Cancer 25:299–305

    Article  CAS  PubMed  Google Scholar 

  26. Chondronasiou D, Martínez de Villarreal J, Melendez E et al (2022) Deciphering the roadmap of in vivo reprogramming toward pluripotency. Stem Cell Rep 17:2501–2517

    Article  CAS  Google Scholar 

  27. Takaishi S, Okumura T, Tu S et al (2009) Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells 27:1006–1020

    Article  CAS  PubMed  Google Scholar 

  28. Niwa H, Ogawa K, Shimosato D, Adachi K (2009) A parallel circuit of LIF signalling pathways maintains pluripotency of mouse ES cells. Nature 460:118–122

    Article  CAS  PubMed  Google Scholar 

  29. Boyd LNC, Andini KD, Peters GJ et al (2022) Heterogeneity and plasticity of cancer-associated fibroblasts in the pancreatic tumor microenvironment. Semin Cancer Biol 82:184–196

    Article  CAS  PubMed  Google Scholar 

  30. Zheng S, Hu C, Lin H et al (2022) circCUL2 induces an inflammatory CAF phenotype in pancreatic ductal adenocarcinoma via the activation of the MyD88-dependent NF-κB signaling pathway. J Experimental Clin Cancer Res 41:1–23

    Article  Google Scholar 

  31. Mello AM, Ngodup T, Lee Y et al (2022) Hypoxia promotes an inflammatory phenotype of fibroblasts in pancreatic cancer. Oncogenesis 11:56

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Schwoerer S, Cimino FV, Ros M et al (2023) Hypoxia potentiates the inflammatory fibroblast phenotype promoted by pancreatic cancer cell-derived cytokines. Cancer Res 83:1596–1610

    Article  Google Scholar 

  33. Vaish U, Jain T, Are AC, Dudeja V (2021) Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma: an update on heterogeneity and therapeutic targeting. Int J Mol Sci 22:13408

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Maeda K, Enomoto A, Hara A, Asai N, Kobayashi T, Horinouchi A et al (2016) Identification of Meflin as a potential marker for mesenchymal stromal cells. Sci Rep 6:1–15

    Article  Google Scholar 

  35. Urbaìn N, Cheung TH (2021) Stem cell quiescence: the challenging path to activation. Dev (Cambridge) 148:dev165084

    Article  Google Scholar 

  36. Prado MM, Frampton AE, Stebbing J, Krell J (2015) Gene of the month: NANOG. J Clin Pathol 68:763–765

    Article  CAS  Google Scholar 

  37. Lu X, Mazur SJ, Lin T, Appella E, Xu Y (2014) The pluripotency factor nanog promotes breast cancer tumorigenesis and metastasis. Oncogene 33:2655–2664

    Article  CAS  PubMed  Google Scholar 

  38. Almeida GM, Pereira C, Park J-H et al (2021) CD44v6 High Membranous expression is a predictive marker of Therapy Response in Gastric Cancer patients. Biomedicines 9:1249

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Bourguignon LYW, Peyrollier K, Xia W, Gilad E (2008) Hyaluronan-CD44 interaction activates stem cell marker nanog, Stat-3-mediated MDR1 gene expression, and ankyrin-regulated multidrug efflux in breast and ovarian tumor cells. J Biol Chem 283:17635–17651

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Numakura S, Uozaki H, Kikuchi Y, Watabe S, Togashi A, Watanabe M (2019) Mesenchymal stem cell marker expression in gastric cancer stroma. Anticancer Res 39:387–393

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

YS-1 cells were donated by Dr. Yutaka Shimada (Department of Nanobio Drug Discovery Graduate School of Pharmaceutical Sciences, Kyoto University) via the JCRB Cell Bank (https://cellbank.nibiohn.go.jp/). DNA Chip Research Inc. acquired mRNA-seq read count data from frozen cultured cells. This research was funded by the Japan Society for the Promotion of Science (JSPS) KAKENHI, grant number 20K16203.

Funding

This research was funded by the Japan Society for the Promotion of Science (JSPS) KAKENHI, grant number 20K16203.

Author information

Authors and Affiliations

  1. Department of Pathology, Teikyo University School of Medicine, 2-11-1 Kaga, Itabashi-ku, Tokyo, 173-8605, Japan

    Satoe Numakura, Masahiro Kato & Hiroshi Uozaki

Authors
  1. Satoe Numakura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Masahiro Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hiroshi Uozaki
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

S.N and H.U contributed to the study conception. S.N and M.K contributed investigation. S.N, M.K and H.U contributed validation. S.N contributed the project administration, funding acquision, methodology, data curation, formal analysis, visualization and writing original draft preparation. H.U reviewed and edited the draft and supervised the study.

Corresponding author

Correspondence to Satoe Numakura.

Ethics declarations

Ethical approval

Human cell lines were obtained from a donor and providers who have adequate ethical considerations.

Consent to participate

Not applicable.

Consent to publish

Not applicable.

Competing interests

The authors have no relevant financial or non-financial interests to disclose.

Additional information

Publisher’s Note

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

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Numakura, S., Kato, M. & Uozaki, H. Discovery of YS-1 as a cell line of gastric inflammatory cancer-associated fibroblasts. Mol Biol Rep 51, 542 (2024). https://doi.org/10.1007/s11033-024-09442-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11033-024-09442-4

Keywords

Search

Navigation