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Thyroid cancer stem-like cell exosomes: regulation of EMT via transfer of lncRNAs

Laboratory Investigationvolume 98pages11331142 (2018) | Download Citation


Thyroid cancers are the most common endocrine malignancy and approximately 2% of thyroid cancers are anaplastic thyroid carcinoma (ATC), one of the most lethal and treatment resistant human cancers. Cancer stem-like cells (CSCs) may initiate tumorigenesis, induce resistance to chemotherapy and radiation therapy, have multipotent capability and may be responsible for recurrent and metastatic disease. The production of CSCs has been linked to epithelial-mesenchymal transition (EMT) and the acquisition of stemness. Exosomes are small (30–150 nm) membranous vesicles secreted by most cells that play a significant role in cell-to-cell communication. Many non-coding RNAs (ncRNA), such as long-non-coding RNAs (lncRNA), can initiate tumorigenesis and the EMT process. Exosomes carry ncRNAs to local and distant cell populations. This study examines secreted exosomes from two in vitro cell culture models; an EMT model and a CSC model. The EMT was induced in a papillary thyroid carcinoma (PTC) cell line by TGFβ1 treatment. Exosomes from this model were isolated and cultured with naïve PTC cells and examined for EMT induction. In the CSC model, exosomes were isolated from a CSC clonal line, cultured with a normal thyroid cell line and examined for EMT induction. The EMT exosomes transferred the lncRNA MALAT1 and EMT effectors SLUG and SOX2; however, EMT was not induced in this model. The exosomes from the CSC model also transferred the lncRNA MALAT1 and the transcription factors SLUG and SOX2 but additionally transferred linc-ROR and induced EMT in the normal thyroid cells. Preliminary siRNA studies directed towards linc-ROR reduced invasion. We hypothesize that CSC exosomes transfer lncRNAs, importantly linc-ROR, to induce EMT and inculcate the local tumor microenvironment and the distant metastatic niche. Therapies directed towards CSCs, their exosomes and/or the lncRNAs they carry may reduce a tumor’s metastatic capacity.

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  1. 1.

    Davies L, Welch HG. Increasing Incidence of Thyroid Cancer in the United States, 1973-2002. JAMA. 2006;295:2164.

  2. 2.

    Sipos JA, Mazzaferri EL. Thyroid cancer epidemiology and prognostic variables. Clin Oncol. 2010;22:395–404.

  3. 3.

    Kondo T, Ezzat S, Asa SL. Pathogenetic mechanisms in thyroid follicular-cell neoplasia. Nat Rev Cancer. 2006;6:292–306.

  4. 4.

    Albores-Saavedra J, Hernandez M, Sanchez-Sosa S, et al. Histologic variants of papillary and follicular carcinomas associated with anaplastic spindle and giant cell carcinomas of the thyroid: an analysis of rhabdoid and thyroglobulin inclusions. Am J Surg Pathol. 2007;31:729–36.

  5. 5.

    Hardin H, Montemayor-Garcia C, Lloyd RV. Thyroid cancer stem-like cells and epithelial-mesenchymal transition in thyroid cancers. Hum Pathol. 2013;44:1707–13.

  6. 6.

    Lloyd RV, Hardin H, Montemayor-Garcia C, et al. Stem cells and cancer stem-like cells in endocrine tissues. Endocr Pathol. 2013;24:1–10.

  7. 7.

    Takano T. Fetal cell carcinogenesis of the thyroid: a modified theory based on recent evidence. Endocr J. 2014;61:311–20.

  8. 8.

    Hardin H, Zhang R, Helein H, et al. The evolving concept of cancer stem-like cells in thyroid cancer and other solid tumors. Lab Invest. 2017;97:1142–51.

  9. 9.

    Théry C, Zitvogel L, Amigorena S. Exosomes: composition, biogenesis and function. Nat Rev Immunol. 2002;2:569–79.

  10. 10.

    Park JE, Tan HS, Datta A, et al. Hypoxic tumor cell modulates its microenvironment to enhance angiogenic and metastatic potential by secretion of proteins and exosomes. Mol Cell Proteom. 2010;9:61085–99.

  11. 11.

    Suetsugu A, Honma K, Saji S, et al. Imaging exosome transfer from breast cancer cells to stroma at metastatic sites in orthotopic nude-mouse models. Adv Drug Deliv Rev. 2013;65:383–90.

  12. 12.

    Roma-Rodrigues C, Fernandes AR, Baptista PV. Exosome in tumour microenvironment: overview of the crosstalk between normal and cancer cells. Biomed Res Int. 2014;2014:179486.

  13. 13.

    Costa-Silva B, Aiello NM, Ocean AJ, et al. Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver. Nat Cell Biol. 2015;17:816–26.

  14. 14.

    Greening DW, Gopal SK, Mathias RA, et al. Emerging roles of exosomes during epithelial-mesenchymal transition and cancer progression. Semin Cell Dev Biol. 2015;40:60–71.

  15. 15.

    Minciacchi VR, Freeman MR, Di Vizio D. Extracellular vesicles in cancer: exosomes, microvesicles and the emerging role of large oncosomes. Semin Cell Dev Biol. 2015;40:41–51.

  16. 16.

    Syn N, Wang L, Sethi G, et al. Exosome-mediated metastasis: from epithelial-mesenchymal transition to escape from immunosurveillance. Trends Pharmacol Sci. 2016;37:606–17.

  17. 17.

    Lee JC, Zhao JT, Gundara J, et al. Papillary thyroid cancer-derived exosomes contain miRNA-146b and miRNA-222. J Surg Res. 2015;196:39–48.

  18. 18.

    Hoshino A, Costa-Silva B, Shen TL, et al. Tumour exosome integrins determine organotropic metastasis. Nature. 2015;527:329–35.

  19. 19.

    Blondal T, Thomsen AR, Krummheuer J, et al. Exosomal microRNA in cell-free urine samples as a source for liquid prostate cancer biopsy. Cancer Res. 2015;75:3987–87.

  20. 20.

    Brock G, Castellanos-Rizaldos E, Hu L, et al. Liquid biopsy for cancer screening, patient stratification and monitoring. Transl Cancer Res. 2015;4:280–90.

  21. 21.

    Hurley J, O’Neill V, Brock G, et al. Abstract 4969: exosomal RNA based liquid biopsy detection of androgen receptor variant 7 in plasma from prostate cancer patients. Cancer Res. 2016;76:4969–69.

  22. 22.

    Barile L, Vassalli G. Exosomes: therapy delivery tools and biomarkers of diseases. Pharmacol Ther. 2017;174:63–78.

  23. 23.

    Li W, Li C, Zhou T, et al. Role of exosomal proteins in cancer diagnosis. Mol Cancer. 2017;16:145.

  24. 24.

    Wang J, Zheng Y, Zhao M. Exosome-based cancer therapy: implication for targeting cancer stem cells. Front Pharmacol. 2016;7:533.

  25. 25.

    Wu CY, Du SL, Zhang J, et al. Exosomes and breast cancer: a comprehensive review of novel therapeutic strategies from diagnosis to treatment. Cancer Gene Ther. 2017;24:6–12.

  26. 26.

    Medema JP. Cancer stem cells: the challenges ahead. Nat Cell Biol. 2013;15:338–44.

  27. 27.

    Craig AJ, von Felden J, Villanueva A. Molecular profiling of liver cancer heterogeneity. Discov Med. 2017;24:117–25.

  28. 28.

    Mani SA, Guo W, Liao MJ, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 2008;133:704–15.

  29. 29.

    Hardin H, Guo Z, Shan W, et al. The roles of the epithelial-mesenchymal transition marker PRRX1 and miR-146b-5p in papillary thyroid carcinoma progression. Am J Pathol. 2014;184:2342–54.

  30. 30.

    Fabregat I, Malfettone A, Soukupova J. New insights into the crossroads between EMT and stemness in the context of cancer. J Clin Med. 2016;5:37.

  31. 31.

    Pradella D, Naro C, Sette C, et al. EMT and stemness: flexible processes tuned by alternative splicing in development and cancer progression. Mol Cancer BioMed Cent. 2017;16:8.

  32. 32.

    Buehler D, Hardin H, Shan W, et al. Expression of epithelial-mesenchymal transition regulators SNAI2 and TWIST1 in thyroid carcinomas. Mod Pathol. 2013;26:54–61.

  33. 33.

    Montemayor-Garcia C, Hardin H, Guo Z, et al. The role of epithelial mesenchymal transition markers in thyroid carcinoma progression. Endocr Pathol. 2013;24:1–14.

  34. 34.

    Heery R, Finn SP, Cuffe S, et al. Long non-coding RNAs: key regulators of epithelial-mesenchymal transition, tumour drug resistance and cancer stem cells. Cancers. 2017;9:38.

  35. 35.

    Rinn JL, Kertesz M, Wang JK, et al. Functional demarcation of active and silent chromatin domains in human HOX loci by noncoding RNAs. Cell. 2007;129:1311–23.

  36. 36.

    Sui F, Ji M, Hou P. Long non-coding RNAs in thyroid cancer: biological functions and clinical significance. Mol Cell Endocrinol. 2017. (in press).

  37. 37.

    Zhang R, Hardin H, Huang W, et al. MALAT1 long non-coding RNA expression in thyroid tissues: analysis by in situ hybridization and real-time PCR. Endocr Pathol. 2017;28:7–12.

  38. 38.

    Weidle UH, Birzele F, Kollmorgen G, et al. Long non-coding RNAs and their role in metastasis. Cancer Genom Proteom. 2017;14:143–60.

  39. 39.

    Hardin H, Yu XM, Harrison AD, et al. Generation of novel thyroid cancer stem-like cell clones effects of resveratrol and valproic acid. Am J Pathol. 2016;186:1662–73.

  40. 40.

    Fierabracci A. Identifying thyroid stem/progenitor cells: advances and limitations. J Endocrinol. 2012;213:1–13.

  41. 41.

    Grelet S, McShane A, Geslain R, et al. Pleiotropic roles of non-coding RNAs in TGF-β-mediated epithelial-mesenchymal transition and their functions in tumor progression. Cancers. 2017;9:75.

  42. 42.

    Brinckerhoff CE. Matrix metalloproteinases in health and disease: sculpting the human body. 1st ed. (Republic of Singapore): World Scientific Publishing Co; 2017.

  43. 43.

    Weidle UH, Birzele F, Kollmorgen G, et al. The multiple roles of exosomes in metastasis. Cancer Genom Proteom. 2017;14:1–16.

  44. 44.

    Zhang R, Hardin H, Wei H, Buehler D, Lloyd RV. Long non-coding RNA linc-ROR is upregulated in papillary thyroid carcinoma. Endocr Pathol. 2017;29:1–8.

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We kindly thank Dr. John A. Copland III (Mayo Clinic, Jacksonville, FL) for the THJ-16T cell line, Dr. Daniel T. Ruan (Brigham and Women’s Hospital, Boston, MA) for the TPC-1 cell line and the staffs of the Flow Cytometry and the 3P laboratories (University of Wisconsin Carbone Cancer Center, Cancer Center Support Grant P30 CA014520) for their services. This work was supported in part by the use of the Electron Microscopy facility at the William S. Middleton Memorial Veterans Hospital, Madison WI. We acknowledge the technical assistance of Joan Sempf and Traci Niesen.


This study was supported by a grant from the University of Wisconsin Carbone Cancer Center (RVL), and by NIH Cancer Center Support Grant P30 CA014520-39 (University of Wisconsin Cancer Center).

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  1. Department of Pathology and Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI, 53792, USA

    • Heather Hardin
    • , Holly Helein
    • , Kristy Meyer
    • , Samantha Robertson
    • , Ranran Zhang
    • , Weixiong Zhong
    •  & Ricardo V. Lloyd


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The authors declare that they have no conflict of interest.

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Correspondence to Ricardo V. Lloyd.

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