Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Loss of exosomal miR-200b-3p from hypoxia cancer-associated fibroblasts promotes tumorigenesis and reduces sensitivity to 5-Flourouracil in colorectal cancer via upregulation of ZEB1 and E2F3

Cancer Gene Therapy (2023)Cite this article

Subjects

Abstract

Hypoxia-mediated tumor progression is a major clinical challenge in human cancers including colorectal cancer (CRC). In addition, exosome-mediated transfer of miRNAs from cancer-associated fibroblasts (CAFs) to cancer cells could promote tumor progression. However, the mechanisms by which hypoxia CAFs promotes CRC progression remain largely unknown. CAFs and normal fibroblasts (NFs) were isolated from CRC tissues and adjacent normal tissues. Next, exosomes were isolated from the supernatant of CAFs that cultured under normoxia (CAFs-N-Exo) and hypoxia (CAFs-H-Exo). RNA-sequencing was then performed to identify differentially expressed miRNAs (DEMs) between CAFs-N-Exo and CAFs-H-Exo. Compared with exosomes derived from normoxia CAFs, exosomes derived from hypoxic CAFs were able to promote CRC cell proliferation, migration, invasion, stemness and reduce the sensitivity of CRC cells to 5-fluorouracil (5-FU). In addition, miR-200b-3p levels were dramatically decreased in exosomes derived from hypoxic CAFs. Remarkably, increasing exosomal miR-200b-3p in hypoxic CAFs reversed the promoting effects of hypoxic CAFs on CRC cell growth in vitro and in vivo. Furthermore, miR-200b-3p agomir could inhibit CRC cell migration, invasion, stemness and increase the sensitivity of SW480 cells to 5-FU via downregulating ZEB1 and E2F3. Collectively, loss of exosomal miR-200b-3p in hypoxia CAFs could contribute to CRC progression via upregulation of ZEB1 and E2F3. Thus, increasing exosomal miR-200b-3p might serve as an alternative approach for the treatment of CRC.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Identification of DEMs between CAFs-N-Exo and CAFs-H-Exo.
Fig. 2: HIF-1α inhibited miR-200b-3p level in hypoxic CAFs.
Fig. 3: Hypoxic CAFs exosomes transferred miR-200b-3p into SW480 cells.
Fig. 4: Loss of miR-200b-3p in hypoxic CAFs-derived exosomes promoted SW480 cell proliferation, migration, invasion.
Fig. 5: Loss of miR-200b-3p in hypoxic CAFs-derived exosomes promoted SW480 cell stemness and reduced the sensitivity of CRC cells to 5-FU.
Fig. 6: ZEB1 and E2F3 were direct targets of exosomal miR-200b-3p in CRC.
Fig. 7: MiR-200b-3p inhibited SW480 cell proliferation, migration, invasion, stemness and increased the sensitivity of SW480 cells to 5-FU via targeting ZEB1 and E2F3.
Fig. 8: Loss of miR-200b-3p in hypoxic CAFs-derived exosomes promoted SW480 cell growth and stemness in vivo.

Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. Balacescu O, Sur D, Cainap C, Visan S, Cruceriu D, Manzat-Saplacan R. The Impact of miRNA in Colorectal Cancer Progression and Its Liver Metastases. Int J Mol Sci. 2018;19:3711.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Wang L, Cho KB, Li Y, Tao G, Xie Z, Guo B. Long Noncoding RNA (lncRNA)-Mediated Competing Endogenous RNA Networks Provide Novel Potential Biomarkers and Therapeutic Targets for Colorectal Cancer. Int J Mol Sci. 2019;20:5758.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Piawah S, Venook AP. Targeted therapy for colorectal cancer metastases: A review of current methods of molecularly targeted therapy and the use of tumor biomarkers in the treatment of metastatic colorectal cancer. Cancer. 2019;125:4139–47.

    Article  PubMed  Google Scholar 

  4. Kuipers EJ, Grady WM, Lieberman D, Seufferlein T, Sung JJ, Boelens PG, et al. Colorectal cancer. Nat Rev Dis Prim. 2015;1:15065.

    Article  PubMed  Google Scholar 

  5. Bashir B, Snook AE. Immunotherapy regimens for metastatic colorectal carcinomas. Hum Vaccin Immunother. 2018;14:250–4.

    Article  PubMed  Google Scholar 

  6. Sousa-Squiavinato ACM, Vasconcelos RI, Gehren AS, Fernandes PV, de Oliveira IM, Boroni M, et al. Cofilin-1, LIMK1 and SSH1 are differentially expressed in locally advanced colorectal cancer and according to consensus molecular subtypes. Cancer cell Int. 2021;21:69.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Aguirre-Portolés C, Feliu J, Reglero G, Ramírez de Molina A. ABCA1 overexpression worsens colorectal cancer prognosis by facilitating tumour growth and caveolin-1-dependent invasiveness, and these effects can be ameliorated using the BET inhibitor apabetalone. Mol Oncol. 2018;12:1735–52.

    Article  PubMed  PubMed Central  Google Scholar 

  8. McKeown SR. Defining normoxia, physoxia and hypoxia in tumours-implications for treatment response. Br J Radiol. 2014;87:20130676.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Krishnamachary B, Mironchik Y, Jacob D, Goggins E, Kakkad S, Ofori F, et al. Hypoxia theranostics of a human prostate cancer xenograft and the resulting effects on the tumor microenvironment. Neoplasia. 2020;22:679–88.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Meng W, Hao Y, He C, Li L, Zhu G. Exosome-orchestrated hypoxic tumor microenvironment. Mol Cancer. 2019;18:57.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Silvoniemi A, Suilamo S, Laitinen T, Forsback S, Löyttyniemi E, Vaittinen S, et al. Repeatability of tumour hypoxia imaging using [(18)F]EF5 PET/CT in head and neck cancer. Eur J Nucl Med Mol imaging. 2018;45:161–9.

    Article  CAS  PubMed  Google Scholar 

  12. Qin Y, Liu HJ, Li M, Zhai DH, Tang YH, Yang L, et al. Salidroside improves the hypoxic tumor microenvironment and reverses the drug resistance of platinum drugs via HIF-1α signaling pathway. EBioMedicine. 2018;38:25–36.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Ni J, Wang X, Stojanovic A, Zhang Q, Wincher M, Bühler L, et al. Single-Cell RNA Sequencing of Tumor-Infiltrating NK Cells Reveals that Inhibition of Transcription Factor HIF-1α Unleashes NK Cell Activity. Immunity. 2020;52:1075–1087.

    Article  CAS  PubMed  Google Scholar 

  14. Campbell EJ, Dachs GU, Morrin HR, Davey VC, Robinson BA, Vissers MCM. Activation of the hypoxia pathway in breast cancer tissue and patient survival are inversely associated with tumor ascorbate levels. BMC Cancer. 2019;19:307.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Garg N, Kumar P, Gadhave K, Giri R. The dark proteome of cancer: Intrinsic disorderedness and functionality of HIF-1α along with its interacting proteins. Prog Mol Biol Transl Sci. 2019;166:371–403.

    Article  CAS  PubMed  Google Scholar 

  16. De Francesco EM, Lappano R, Santolla MF, Marsico S, Caruso A, Maggiolini M. HIF-1α/GPER signaling mediates the expression of VEGF induced by hypoxia in breast cancer associated fibroblasts (CAFs). Breast Cancer Res. 2013;15:R64.

    Article  PubMed  PubMed Central  Google Scholar 

  17. Wu HJ, Hao M, Yeo SK, Guan JL. FAK signaling in cancer-associated fibroblasts promotes breast cancer cell migration and metastasis by exosomal miRNAs-mediated intercellular communication. Oncogene. 2020;39:2539–49.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Sazeides C, Le A. Metabolic Relationship between Cancer-Associated Fibroblasts and Cancer Cells. Adv Exp Med Biol. 2018;1063:149–65.

    Article  CAS  PubMed  Google Scholar 

  19. Su S, Chen J, Yao H, Liu J, Yu S, Lao L, et al. CD10(+)GPR77(+) Cancer-Associated Fibroblasts Promote Cancer Formation and Chemoresistance by Sustaining Cancer Stemness. Cell. 2018;172:841–856.e16.

    Article  CAS  PubMed  Google Scholar 

  20. Deep G, Panigrahi GK. Hypoxia-Induced Signaling Promotes Prostate Cancer Progression: Exosomes Role as Messenger of Hypoxic Response in Tumor Microenvironment. Crit Rev Oncog. 2015;20:419–34.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Kugeratski FG, Atkinson SJ, Neilson LJ, Lilla S, Knight JRP, Serneels J. Hypoxic cancer-associated fibroblasts increase NCBP2-AS2/HIAR to promote endothelial sprouting through enhanced VEGF signaling. Sci Signal. 2019;12:eaan8247.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Wang H, Wei H, Wang J, Li L, Chen A, Li Z. MicroRNA-181d-5p-Containing Exosomes Derived from CAFs Promote EMT by Regulating CDX2/HOXA5 in Breast Cancer. Mol Ther Nucleic Acids. 2020;19:654–67.

    Article  CAS  PubMed  Google Scholar 

  23. Ge Q, Zhou Y, Lu J, Bai Y, Xie X, Lu Z. miRNA in plasma exosome is stable under different storage conditions. Molecules. 2014;19:1568–75.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Richards KE, Zeleniak AE, Fishel ML, Wu J, Littlepage LE, Hill R. Cancer-associated fibroblast exosomes regulate survival and proliferation of pancreatic cancer cells. Oncogene. 2017;36:1770–8.

    Article  CAS  PubMed  Google Scholar 

  25. Kalluri R. The biology and function of exosomes in cancer. J Clin Investig. 2016;126:1208–15.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Hu JL, Wang W, Lan XL, Zeng ZC, Liang YS, Yan YR, et al. CAFs secreted exosomes promote metastasis and chemotherapy resistance by enhancing cell stemness and epithelial-mesenchymal transition in colorectal cancer. Mol Cancer. 2019;18:91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Li YY, Zhou CX, Gao Y. Interaction between oral squamous cell carcinoma cells and fibroblasts through TGF-β1 mediated by podoplanin. Exp Cell Res. 2018;369:43–53.

    Article  CAS  PubMed  Google Scholar 

  28. Zeng Z, Li Y, Pan Y, Lan X, Song F, Sun J, et al. Cancer-derived exosomal miR-25-3p promotes pre-metastatic niche formation by inducing vascular permeability and angiogenesis. Nat Commun. 2018;9:5395.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Ritchie ME, Phipson B, Wu D, Hu Y, Law CW, Shi W, et al. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 2015;43:e47.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Wanandi SI, Ningsih SS, Asikin H, Hosea R, Neolaka GMG. Metabolic Interplay between Tumour Cells and Cancer-Associated Fibroblasts (CAFs) under Hypoxia versus Normoxia. Malays J Med Sci. 2018;25:7–16.

    PubMed  PubMed Central  Google Scholar 

  31. Petrova V, Annicchiarico-Petruzzelli M, Melino G, Amelio I. The hypoxic tumour microenvironment. Oncogenesis. 2018;7:10.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Huang GL, Sun J, Lu Y, Liu Y, Cao H, Zhang H, et al. MiR-200 family and cancer: From a meta-analysis view. Mol Asp Med. 2019;70:57–71.

    Article  CAS  Google Scholar 

  33. Cavallari I, Ciccarese F, Sharova E, Urso L, Raimondi V, Silic-Benussi M. et al. The miR-200 Family of microRNAs: Fine Tuners of Epithelial-Mesenchymal Transition and Circulating. Cancer Biomarkers. Cancers. 2021;13:5874.

    PubMed  Google Scholar 

  34. Chen L, Wang X, Zhu Y, Zhu J, Lai Q. miR‑200b‑3p inhibits proliferation and induces apoptosis in colorectal cancer by targeting Wnt1. Mol Med Rep. 2018;18:2571–80.

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Wu J, Cui H, Zhu Z, Wang L. MicroRNA-200b-3p suppresses epithelial-mesenchymal transition and inhibits tumor growth of glioma through down-regulation of ERK5. Biochem Biophys Res Commun. 2016;478:1158–64.

    Article  CAS  PubMed  Google Scholar 

  36. Karshovska E, Wei Y, Subramanian P, Mohibullah R, Geißler C, Baatsch I, et al. HIF-1α (Hypoxia-Inducible Factor-1α) Promotes Macrophage Necroptosis by Regulating miR-210 and miR-383. Arterioscler Thromb Vasc Biol. 2020;40:583–96.

    Article  CAS  PubMed  Google Scholar 

  37. Wu L, Chen Y, Chen Y, Yang W, Han Y, Lu L, et al. Effect of HIF-1α/miR-10b-5p/PTEN on Hypoxia-Induced Cardiomyocyte Apoptosis. J Am Heart Assoc. 2019;8:e011948.

    Article  PubMed  PubMed Central  Google Scholar 

  38. Tutar Y. miRNA and cancer; computational and experimental approaches. Curr Pharm Biotechnol. 2014;15:429.

    Article  CAS  PubMed  Google Scholar 

  39. Ferragut Cardoso AP, Udoh KT, States JC. Arsenic-induced changes in miRNA expression in cancer and other diseases. Toxicol Appl Pharmacol. 2020;409:115306.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Noman MZ, Messai Y, Carré T, Akalay I, Méron M, Janji B, et al. Microenvironmental hypoxia orchestrating the cell stroma cross talk, tumor progression and antitumor response. Crit Rev Immunol. 2011;31:357–77.

    Article  CAS  PubMed  Google Scholar 

  41. Zhao H, Yang L, Baddour J, Achreja A, Bernard V, Moss T, et al. Tumor microenvironment derived exosomes pleiotropically modulate cancer cell metabolism. eLife. 2016;5:e10250.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Monteran L, Erez N. The Dark Side of Fibroblasts: Cancer-Associated Fibroblasts as Mediators of Immunosuppression in the Tumor Microenvironment. Front Immunol. 2019;10:1835.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Yang F, Yan Y, Yang Y, Hong X, Wang M, Yang Z, et al. MiR-210 in exosomes derived from CAFs promotes non-small cell lung cancer migration and invasion through PTEN/PI3K/AKT pathway. Cell Signal. 2020;73:109675.

    Article  CAS  PubMed  Google Scholar 

  44. Li YY, Tao YW, Gao S, Li P, Zheng JM, Zhang SE, et al. Cancer-associated fibroblasts contribute to oral cancer cells proliferation and metastasis via exosome-mediated paracrine miR-34a-5p. EBioMedicine. 2018;36:209–20.

    Article  PubMed  PubMed Central  Google Scholar 

  45. Xia X, Wang S, Ni B, Xing S, Cao H, Zhang Z, et al. Hypoxic gastric cancer-derived exosomes promote progression and metastasis via MiR-301a-3p/PHD3/HIF-1α positive feedback loop. Oncogene. 2020;39:6231–44.

    Article  CAS  PubMed  Google Scholar 

  46. Qin X, Guo H, Wang X, Zhu X, Yan M, Wang X, et al. Exosomal miR-196a derived from cancer-associated fibroblasts confers cisplatin resistance in head and neck cancer through targeting CDKN1B and ING5. Genome Biol. 2019;20:12.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Richard G, Dalle S, Monet MA, Ligier M, Boespflug A, Pommier RM, et al. ZEB1-mediated melanoma cell plasticity enhances resistance to MAPK inhibitors. EMBO Mol Med. 2016;8:1143–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Cortés M, Sanchez-Moral L, de Barrios O, Fernández-Aceñero MJ, Martínez-Campanario MC, Esteve-Codina A, et al. Tumor-associated macrophages (TAMs) depend on ZEB1 for their cancer-promoting roles. EMBO J. 2017;36:3336–55.

    Article  PubMed  PubMed Central  Google Scholar 

  49. Goulet CR, Champagne A, Bernard G, Vandal D, Chabaud S, Pouliot F, et al. Cancer-associated fibroblasts induce epithelial-mesenchymal transition of bladder cancer cells through paracrine IL-6 signalling. BMC Cancer. 2019;19:137.

    Article  PubMed  PubMed Central  Google Scholar 

  50. Ma M, Wang X, Chen X, Cai R, Chen F, Dong W, et al. MicroRNA-432 targeting E2F3 and P55PIK inhibits myogenesis through PI3K/AKT/mTOR signaling pathway. RNA Biol. 2017;14:347–60.

    Article  PubMed  PubMed Central  Google Scholar 

  51. Jusino S, Rivera-Rivera Y, Chardón-Colón C, Ruiz-Justiz AJ, Vélez-Velázquez J, Isidro A, et al. E2F3 drives the epithelial-to-mesenchymal transition, cell invasion, and metastasis in breast cancer. Exp Biol Med. 2021;246:2057–71.

    Article  CAS  Google Scholar 

  52. Wu QB, Sheng X, Zhang N, Yang MW, Wang F. Role of microRNAs in the resistance of colorectal cancer to chemoradiotherapy. Mol Clin Oncol. 2018;8:523–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  53. Feng B, Wang R, Song HZ, Chen LB. MicroRNA-200b reverses chemoresistance of docetaxel-resistant human lung adenocarcinoma cells by targeting E2F3. Cancer. 2012;118:3365–76.

    Article  CAS  PubMed  Google Scholar 

  54. Voissiere A, Jouberton E, Maubert E, Degoul F, Peyrode C, Chezal JM, et al. Development and characterization of a human three-dimensional chondrosarcoma culture for in vitro drug testing. PloS One. 2017;12:e0181340.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Funding

The present study was financially supported by the Natural Science Foundation of Zhejiang Province (LY18H160041, LY17H160064, LQ21H160042), the Funding Project of Health and Family Planning Commission of Zhejiang Province (2018KY217), and the Funding Project Administration of Traditional Chinese Medicine of Zhejiang Province (2018ZA009).

Author information

Author notes

  1. These authors contributed equally: Wenjing Gong, Yang Guo, Hang Yuan.

Authors and Affiliations

  1. General Surgery, Cancer Center, Department of Colorectal Surgery, Zhejiang Provincial People’s Hospital (Affiliated People’s Hospital, Hangzhou Medical College), Hangzhou, Zhejiang, 310014, PR China

    Wenjing Gong, Yang Guo, Hang Yuan, Rui Chai, Ziang Wan, Boan Zheng, Xinye Hu, Bingchen Chen, Shan Gao, Qiaoqiong Dai, Peng Yu & Shiliang Tu

Authors
  1. Wenjing Gong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yang Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hang Yuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rui Chai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ziang Wan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Boan Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xinye Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Bingchen Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Shan Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Qiaoqiong Dai
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Peng Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Shiliang Tu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

WG, YG and HY carried out experiments and made major contributions to the design and manuscript drafting of this study. RC, ZW, BZ, XH, BC, SG, QD and PY were responsible for investigation, data analysis, data interpretation. ST conceived experiments and revised the manuscript critically for important intellectual content. All authors agreed to be accountable for all aspects of the work. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Shiliang Tu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

All procedures were approved by the ethics committee of the Zhejiang Provincial People’s Hospital.

Additional information

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

Supplementary information

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

Gong, W., Guo, Y., Yuan, H. et al. Loss of exosomal miR-200b-3p from hypoxia cancer-associated fibroblasts promotes tumorigenesis and reduces sensitivity to 5-Flourouracil in colorectal cancer via upregulation of ZEB1 and E2F3. Cancer Gene Ther (2023). https://doi.org/10.1038/s41417-023-00591-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1038/s41417-023-00591-5

Search

Advanced search

Quick links