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.

  • Article
  • Published:

Msi1 promotes breast cancer metastasis by regulating invadopodia-mediated extracellular matrix degradation via the Timp3–Mmp9 pathway

Oncogene volume 40, pages 4832–4845 (2021)Cite this article

Subjects

Abstract

Metastasis is the main cause of death in breast cancer patients. The initial step of metastasis is invadopodia-mediated extracellular matrix (ECM) degradation, which enables local breast tumor cells to invade surrounding tissues. However, the molecular mechanism underlying invadopodia-mediated metastasis remains largely unknown. Here we found that the RNA-binding protein Musashi1 (Msi1) exhibited elevated expression in invasive breast tumors and promoted lung metastasis of mammary cancer cells. Suppression of Msi1 reduced invadopodia formation in mammary cancer cells. Furthermore, Msi1 deficiency decreased the expression and activity of Mmp9, an important enzyme in ECM degradation. Mechanistically, Msi1 directly suppressed Timp3, an endogenous inhibitor of Mmp9. In clinical breast cancer specimens, TIMP3 and MSI1 levels were significantly inversely correlated both in normal breast tissue and breast cancer tissues and associated with overall survival in breast cancer patients. Taken together, our findings demonstrate that the MSI1–TIMP3–MMP9 cascade is critical for invadopodia-mediated onset of metastasis in breast cancer, providing novel insights into a promising therapeutic strategy for breast cancer metastasis.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Fig. 1: MSI1 expression is upregulated in invasive breast cancer.
Fig. 2: Msi1 depletion impairs lung metastasis of mammary tumor cells rather than tumor growth.
Fig. 3: Msi1 depletion reduces mammary tumor cell invasion and migration.
Fig. 4: Msi1 deficiency blocks invadopodia formation and inhibits Mmp9 expression and activity in mammary tumor cells.
Fig. 5: Msi1 directly suppresses Timp3 to regulate ECM degradation.
Fig. 6: Timp3 functionally mediated the promotive effects of Msi1 on breast cancer cell invasion and lung metastasis.
Fig. 7: TIMP3 expression is inversely correlated with MSI1 expression in human breast cancer.
Fig. 8: Schematic of the function and mechanism of Msi1 in breast cancer metastasis.

Similar content being viewed by others

References

  1. Weigelt B, Peterse JL, van ‘t Veer LJ. Breast cancer metastasis: markers and models. Nat Rev Cancer. 2005;5:591–602.

    Article  CAS  PubMed  Google Scholar 

  2. Chiang AC, Massague J. Molecular basis of metastasis. N. Engl J Med. 2008;359:2814–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Carman CV, Sage PT, Sciuto TE, de la Fuente MA, Geha RS, Ochs HD, et al. Transcellular diapedesis is initiated by invasive podosomes. Immunity. 2007;26:784–97.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Clark ES, Whigham AS, Yarbrough WG, Weaver AM. Cortactin is an essential regulator of matrix metalloproteinase secretion and extracellular matrix degradation in invadopodia. Cancer Res. 2007;67:4227–35.

    Article  CAS  PubMed  Google Scholar 

  5. Oser M, Yamaguchi H, Mader CC, Bravo-Cordero JJ, Arias M, Chen X, et al. Cortactin regulates cofilin and N-WASp activities to control the stages of invadopodium assembly and maturation. J Cell Biol. 2009;186:571–87.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Paz H, Pathak N, Yang J. Invading one step at a time: the role of invadopodia in tumor metastasis. Oncogene. 2014;33:4193–202.

    Article  CAS  PubMed  Google Scholar 

  7. Poincloux R, Lizarraga F, Chavrier P. Matrix invasion by tumour cells: a focus on MT1-MMP trafficking to invadopodia. J Cell Sci. 2009;122:3015–24.

    Article  CAS  PubMed  Google Scholar 

  8. Bourguignon LY, Gunja-Smith Z, Iida N, Zhu HB, Young LJ, Muller WJ, et al. CD44v(3,8-10) is involved in cytoskeleton-mediated tumor cell migration and matrix metalloproteinase (MMP-9) association in metastatic breast cancer cells. J Cell Physiol. 1998;176:206–15.

    Article  CAS  PubMed  Google Scholar 

  9. Jacob A, Prekeris R. The regulation of MMP targeting to invadopodia during cancer metastasis. Front Cell Dev Biol. 2015;3:4.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Ala-aho R, Kähäri V-M. Collagenases in cancer. Biochimie. 2005;87:273–86.

    Article  CAS  PubMed  Google Scholar 

  11. Mehner C, Hockla A, Miller E, Ran S, Radisky DC, Radisky ES. Tumor cell-produced matrix metalloproteinase 9 (MMP-9) drives malignant progression and metastasis of basal-like triple negative breast cancer. Oncotarget. 2014;5:2736–49.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Gomez DE, Alonso DF, Yoshiji H, Thorgeirsson UP. Tissue inhibitors of metalloproteinases: structure, regulation and biological functions. Eur J Cell Biol. 1997;74:111–22.

    CAS  PubMed  Google Scholar 

  13. Brew K, Nagase H. The tissue inhibitors of metalloproteinases (TIMPs): an ancient family with structural and functional diversity. Biochim Biophys Acta. 2010;1803:55–71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Pavloff N, Staskus PW, Kishnani NS, Hawkes S. A new inhibitor of metalloproteinases from chicken: ChIMP-3. A third member of the TIMP family. J Biol Chem. 1992;267:17321–6.

    Article  CAS  PubMed  Google Scholar 

  15. Anand-Apte B, Bao L, Smith R, Iwata K, Olsen BR, Zetter B, et al. A review of tissue inhibitor of metalloproteinases-3 (TIMP-3) and experimental analysis of its effect on primary tumor growth. Biochem Cell Biol. 1996;74:853–62.

    Article  CAS  PubMed  Google Scholar 

  16. Zhang Z, Wang J, Wang X, Song W, Shi Y, Zhang L. MicroRNA-21 promotes proliferation, migration, and invasion of cervical cancer through targeting TIMP3. Arch Gynecol Obstet. 2018;297:433–42.

    Article  CAS  PubMed  Google Scholar 

  17. Olivares-Urbano MA, Grinan-Lison C, Zurita M, Del Moral R, Rios-Arrabal S, Artacho-Cordon F, et al. Matrix metalloproteases and TIMPs as prognostic biomarkers in breast cancer patients treated with radiotherapy: a pilot study. J Cell Mol Med. 2020;24:139–48.

    Article  CAS  PubMed  Google Scholar 

  18. Li W, Yi J, Zheng X, Liu S, Fu W, Ren L, et al. miR-29c plays a suppressive role in breast cancer by targeting the TIMP3/STAT1/FOXO1 pathway. Clin Epigenetics. 2018;10:64.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Vo DT, Qiao M, Smith AD, Burns SC, Brenner AJ, Penalva LO. The oncogenic RNA-binding protein Musashi1 is regulated by tumor suppressor miRNAs. RNA Biol. 2011;8:817–28.

    Article  CAS  PubMed  Google Scholar 

  20. Sanchez-Diaz PC, Burton TL, Burns SC, Hung JY, Penalva LO. Musashi1 modulates cell proliferation genes in the medulloblastoma cell line Daoy. BMC Cancer. 2008;8:280.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Imai T, Tokunaga A, Yoshida T, Hashimoto M, Mikoshiba K, Weinmaster G, et al. The neural RNA-binding protein Musashi1 translationally regulates mammalian numb gene expression by interacting with its mRNA. Mol Cell Biol. 2001;21:3888–900.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Ascano M, Gerstberger S, Tuschl T. Multi-disciplinary methods to define RNA-protein interactions and regulatory networks. Curr Opin Genet Dev. 2013;23:20–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Henras AK, Soudet J, Gerus M, Lebaron S, Caizergues-Ferrer M, Mougin A, et al. The post-transcriptional steps of eukaryotic ribosome biogenesis. Cell Mol Life Sci. 2008;65:2334–59.

    Article  CAS  PubMed  Google Scholar 

  24. de Sousa Abreu R, Sanchez-Diaz PC, Vogel C, Burns SC, Ko D, Burton TL, et al. Genomic analyses of musashi1 downstream targets show a strong association with cancer-related processes. J Biol Chem. 2009;284:12125–35.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Lin J-C, Tsai J-T, Chao T-Y, Ma H-I, Chien C-S, Liu W-H. MSI1 associates glioblastoma radioresistance via homologous recombination repair, tumor invasion and cancer stem-like cell properties. Radiother Oncol. 2018;129:352–63.

    Article  CAS  PubMed  Google Scholar 

  26. Gong P, Wang Y, Gao Y, Gao M, Liu L, Qu P, et al. Msi1 promotes tumor progression by epithelial-to-mesenchymal transition in cervical cancer. Hum Pathol. 2017;65:53–61.

    Article  CAS  PubMed  Google Scholar 

  27. Wang X-Y, Penalva LOF, Yuan H, Linnoila RI, Lu J, Okano H, et al. Musashi1 regulates breast tumor cell proliferation and is a prognostic indicator of poor survival. Mol Cancer. 2010;9:221.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Khokha R, Werb Z. Mammary gland reprogramming: metalloproteinases couple form with function. Cold Spring Harb Perspect Biol. 2011;3:a004333.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  29. Guy CT, Cardiff RD, Muller WJ. Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease. Mol Cell Biol. 1992;12:954–61.

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Cheung KJ, Gabrielson E, Werb Z, Ewald AJ. Collective invasion in breast cancer requires a conserved basal epithelial program. Cell. 2013;155:1639–51.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Cheung KJ, Ewald AJ. A collective route to metastasis: seeding by tumor cell clusters. Science. 2016;352:167–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Courtneidge SA, Azucena EF, Pass I, Seals DF, Tesfay L. The SRC substrate Tks5, podosomes (invadopodia), and cancer cell invasion. Cold Spring Harb Symp Quant Biol. 2005;70:167–71.

    Article  CAS  PubMed  Google Scholar 

  33. Eckert MA, Lwin TM, Chang AT, Kim J, Danis E, Ohno-Machado L, et al. Twist1-induced invadopodia formation promotes tumor metastasis. Cancer Cell. 2011;19:372–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Schoumacher M, Goldman RD, Louvard D, Vignjevic DM. Actin, microtubules, and vimentin intermediate filaments cooperate for elongation of invadopodia. J Cell Biol. 2010;189:541–56.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Ayala I, Baldassarre M, Giacchetti G, Caldieri G, Tete S, Luini A, et al. Multiple regulatory inputs converge on cortactin to control invadopodia biogenesis and extracellular matrix degradation. J Cell Sci. 2008;121:369–78.

    Article  CAS  PubMed  Google Scholar 

  36. Okano H, Imai T, Okabe M. Musashi: a translational regulator of cell fate. J Cell Sci. 2002;115:1355–9.

    Article  CAS  PubMed  Google Scholar 

  37. Fox RG, Lytle NK, Jaquish DV, Park FD, Ito T, Bajaj J, et al. Image-based detection and targeting of therapy resistance in pancreatic adenocarcinoma. Nature. 2016;534:407–11.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  38. Chen HY, Lin LT, Wang ML, Laurent B, Hsu CH, Pan CM, et al. Musashi-1 enhances glioblastoma cell migration and cytoskeletal dynamics through translational inhibition of Tensin3. Sci Rep. 2017;7:8710.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  39. Nahas GR, Murthy RG, Patel SA, Ganta T, Greco SJ, Rameshwar P. The RNA-binding protein Musashi 1 stabilizes the oncotachykinin 1 mRNA in breast cancer cells to promote cell growth. FASEB J. 2016;30:149–59.

    Article  CAS  PubMed  Google Scholar 

  40. Katz Y, Li F, Lambert NJ, Sokol ES, Tam WL, Cheng AW, et al. Musashi proteins are post-transcriptional regulators of the epithelial-luminal cell state. Elife. 2014;3:e03915.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Li M, Li AQ, Zhou SL, Lv H, Wei P, Yang WT. RNA-binding protein MSI2 isoforms expression and regulation in progression of triple-negative breast cancer. J Exp Clin Cancer Res. 2020;39:92.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  42. Kang MH, Jeong KJ, Kim WY, Lee HJ, Gong G, Suh N, et al. Musashi RNA-binding protein 2 regulates estrogen receptor 1 function in breast cancer. Oncogene. 2017;36:1745–52.

    Article  CAS  PubMed  Google Scholar 

  43. Wang S, Yuan Y, Liao L, Kuang SQ, Tien JC, O’Malley BW, et al. Disruption of the SRC-1 gene in mice suppresses breast cancer metastasis without affecting primary tumor formation. Proc Natl Acad Sci USA. 2009;106:151–6.

    Article  CAS  PubMed  Google Scholar 

  44. Wang Z, Zhang F, He J, Wu P, Tay LWR, Cai M, et al. Binding of PLD2-generated phosphatidic acid to KIF5B promotes MT1-MMP surface trafficking and lung metastasis of mouse breast cancer cells. Dev Cell. 2017;43:186.e7–97.e7.

    Google Scholar 

  45. Murphy DA, Courtneidge SA. The ‘ins’ and ‘outs’ of podosomes and invadopodia: characteristics, formation and function. Nat Rev Mol Cell Biol. 2011;12:413–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Deryugina EI, Quigley JP. Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev. 2006;25:9–34.

    Article  CAS  PubMed  Google Scholar 

  47. Owyong M, Chou J, van den Bijgaart RJ, Kong N, Efe G, Maynard C, et al. MMP9 modulates the metastatic cascade and immune landscape for breast cancer anti-metastatic therapy. Life Sci. Alliance. 2019;2:e201800226.

  48. Sun Y, Wang X, Zhou Q, Lu Y, Zhang H, Chen Q, et al. Inhibitory effect of emodin on migration, invasion and metastasis of human breast cancer MDA-MB-231 cells in vitro and in vivo. Oncol Rep. 2015;33:338–46.

    Article  CAS  PubMed  Google Scholar 

  49. Aktas B, Tewes M, Fehm T, Hauch S, Kimmig R, Kasimir-Bauer S. Stem cell and epithelial-mesenchymal transition markers are frequently overexpressed in circulating tumor cells of metastatic breast cancer patients. Breast Cancer Res. 2009;11:R46.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  50. Raimondi C, Gradilone A, Naso G, Vincenzi B, Petracca A, Nicolazzo C, et al. Epithelial-mesenchymal transition and stemness features in circulating tumor cells from breast cancer patients. Breast Cancer Res Treat. 2011;130:449–55.

    Article  CAS  PubMed  Google Scholar 

  51. Fehm T, Hoffmann O, Aktas B, Becker S, Solomayer EF, Wallwiener D, et al. Detection and characterization of circulating tumor cells in blood of primary breast cancer patients by RT-PCR and comparison to status of bone marrow disseminated cells. Breast Cancer Res. 2009;11:R59.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  52. Tinhofer I, Saki M, Niehr F, Keilholz U, Budach V. Cancer stem cell characteristics of circulating tumor cells. Int J Radiat Biol. 2014;90:622–7.

    Article  CAS  PubMed  Google Scholar 

  53. Charafe-Jauffret E, Ginestier C, Iovino F, Wicinski J, Cervera N, Finetti P, et al. Breast cancer cell lines contain functional cancer stem cells with metastatic capacity and a distinct molecular signature. Cancer Res. 2009;69:1302–13.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Li N, Yousefi M, Nakauka-Ddamba A, Li F, Vandivier L, Parada K, et al. The Msi family of RNA-binding proteins function redundantly as intestinal oncoproteins. Cell Rep. 2015;13:2440–55.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Castagnino A, Castro-Castro A, Irondelle M, Guichard A, Lodillinsky C, Fuhrmann L, et al. Coronin 1C promotes triple-negative breast cancer invasiveness through regulation of MT1-MMP traffic and invadopodia function. Oncogene. 2018;37:6425–41.

    Article  CAS  PubMed  Google Scholar 

  56. Shi J, Ma X, Su Y, Song Y, Tian Y, Yuan S, et al. MiR-31 mediates inflammatory signaling to promote re-epithelialization during skin wound healing. J Invest Dermatol. 2018;138:2253–63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Artym VV, Yamada KM, Mueller SC. ECM degradation assays for analyzing local cell invasion. Methods Mol Biol. 2009;522:211–9.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work is supported by the National Natural Science Foundation of China (Nos. 81772984, 81572614), Beijing Nature Foundation Grant (5162018), the Major Project for Cultivation Technology (2016ZX08008001, 2014ZX08008001), Basic Research Program (2019TC227, 2019TC088), and SKLAB Open Grant (2020SKLAB6-18). The authors thank the members of the laboratory animal center in China Agricultural University for their assistance in animal care.

Author information

Author notes

  1. These authors contributed equally: Xueyun Bi, Pengbo Lou.

Authors and Affiliations

  1. Key Laboratory of Precision Nutrition and Food Quality, Ministry of Education, Department of Nutrition and Health, College of Biological Sciences, China Agricultural University, Beijing, China

    Xueyun Bi, Pengbo Lou, Xiaole Sheng, Ruiqi Liu, Min Deng, Xu Yang, Guilin Li & Zhengquan Yu

  2. The State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock and Research Center for Animal Genetic Resources of Mongolia Plateau College of Life Sciences, Inner Mongolia University, Hohhot, China

    Yongli Song

  3. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin’s Clinical Research Center for Cancer, Tianjin Medical University Cancer Institute and Hospital, Tianjin Medical University, Tianjin, China

    Shukai Yuan & Li Zhao

  4. Center for Scientific Research, Yunnan University of Chinese Medicine, Kunming, Yunnan, China

    Honglei Zhang

  5. State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, China

    Baowei Jiao

  6. College of Veterinary Medicine, China Agricultural University, Beijing, China

    Bing Zhang

  7. Medical Research Center, Department of Radiation Oncology, Peking University Third Hospital, Beijing, China

    Lixiang Xue

  8. State Key Laboratory of Molecular Oncology, National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences, Beijing, China

    Zhihua Liu

  9. Department of Developmental and Cell Biology, Sue and Bill Gross Stem Cell Research Center, Center for Complex Biological Systems, University of California, Irvine, Irvine, CA, USA

    Maksim V. Plikus

  10. Key Laboratory of Precision Nutrition and Food Quality, Ministry of Education, Department of Nutrition and Health, College of Food Sciences and Nutritional Engineering, China Agricultural University, Beijing, China

    Fazheng Ren

  11. CAS Key Laboratory of Bio-medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu, China

    Shan Gao

Authors
  1. Xueyun Bi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pengbo Lou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yongli Song
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xiaole Sheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ruiqi Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Min Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xu Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Guilin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Shukai Yuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Honglei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Baowei Jiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Bing Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Lixiang Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Zhihua Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Maksim V. Plikus
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Fazheng Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Shan Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Li Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Zhengquan Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Li Zhao or Zhengquan Yu.

Ethics declarations

Competing interests

The authors declare no competing interests.

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

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bi, X., Lou, P., Song, Y. et al. Msi1 promotes breast cancer metastasis by regulating invadopodia-mediated extracellular matrix degradation via the Timp3–Mmp9 pathway. Oncogene 40, 4832–4845 (2021). https://doi.org/10.1038/s41388-021-01873-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-021-01873-8

This article is cited by

Search

Advanced search

Quick links