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Endocrine

, Volume 53, Issue 1, pp 145–156 | Cite as

Molecular profiles of cancer stem-like cell populations in aggressive thyroid cancers

  • Mariavittoria Dima
  • Valeria Pecce
  • Mauro Biffoni
  • Cira Rosaria Tiziana Di Gioia
  • Giovanni Tallini
  • Marco Biffoni
  • Francesca Rosignolo
  • Antonella Verrienti
  • Marialuisa Sponziello
  • Giuseppe Damante
  • Diego RussoEmail author
  • Cosimo Durante
Original Article

Abstract

A substantial proportion of patients with advanced thyroid carcinoma fail to respond to or at some point become refractory to conventional therapies. This resistance and the phenomena of thyroid cancer progression and metastasis themselves are thought to be related to tumor-cell sub-populations with stem-like properties. We isolated thyrospheres from four advanced thyroid carcinomas that were resistant to radioiodine therapy and analyzed their molecular profiles. ALDH activity and proteomic profile of main stem cell markers were used to assess stem cell properties. The TaqMan Low Density Array approach was used to evaluate the expression of several genes involved in the EMT process. The phosphorylation status of tyrosine kinase receptors (RTKs) was analyzed to identify potential markers for targeted therapies. We then investigated the effects of the EMT-inhibitor crizotinib on both cell proliferation and phosphorylation status of RTK targets. The cancer stem-like properties of a subset of cells from primary cultures of each tumor were demonstrated. A wide variability among thyrospheres arising from the four thyroid cancers in terms of ALDH activity, stem cell marker expression, and phosphoproteome profiling was present. Dysregulated expression of genes involved in the EMT was observed in all four thyrosphere lines. Treatment with crizotinib was ineffective in cancer stem-like cells, suggesting the presence of a mechanism of resistance in thyrospheres. Collectively, our data indicate that thyroid cancer stem-like populations vary markedly from tumor to tumor and require detailed molecular and biological characterization if they are to be used as the basis of “personalized” treatment of aggressive disease.

Keywords

Metastatic thyroid cancer Epithelial-mesenchymal transition Cancer stem cells Drug resistance 

Notes

Acknowledgments

This study was funded by grants from the Italian Ministry of Universities and Research (FIRB 2008—number RBFR082XL7_003 to C.D., MIUR/Cofin 2010-2011 to D.R.) and from the Fondazione Umberto Di Mario ONLUS. The manuscript was edited by Marian Everett Kent, BSN.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.

Supplementary material

12020_2015_739_MOESM1_ESM.tif (989 kb)
Supplementary Figure 1 Identification of mutations by Sanger Sequencing analysis. NRASQ61K (left) and BRAFV600E (right) mutations detected in tissue specimens 2 and 3 and their corresponding primary tumor cell lines, respectively. Analysis has been performed by Sanger Sequencing of exon 3 of NRAS and exon 15 of BRAF (TIFF 988 kb)
12020_2015_739_MOESM2_ESM.tif (1.9 mb)
Supplementary Figure 2 EMT-related gene expression level in thyrospheres and primary cell populations from all 4 tumors. Columns (error bars) represent means (± SD) of thyrospheres tested with TLDA. Dashed-line represents primary cells (calibrator, set at 1). Differences between thyrosphere expression levels and those of corresponding primary cells were assessed with the Student t-test (* p<0.05; ** p<0.01; *** p<0.001) (TIFF 1929 kb)
12020_2015_739_MOESM3_ESM.tif (1.4 mb)
Supplementary Figure 3 mRNA levels of thyroid-specific genes in thyrospheres and primary cell populations from all 4 tumors. Columns (error bars) represent means (± SD) of 2-ΔCt values in thyrosphere and primary cell populations tested with TLDA. Differences between thyrosphere expression levels and those of corresponding primary cells (calibrator) were assessed with the Student t-test (* p<0.05; ** p<0.01; *** p<0.001) (TIFF 1420 kb)
12020_2015_739_MOESM4_ESM.pdf (64 kb)
Supplementary material 4 (PDF 64 kb)
12020_2015_739_MOESM5_ESM.pdf (116 kb)
Supplementary material 5 (PDF 115 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Mariavittoria Dima
    • 1
  • Valeria Pecce
    • 1
  • Mauro Biffoni
    • 2
  • Cira Rosaria Tiziana Di Gioia
    • 3
  • Giovanni Tallini
    • 4
  • Marco Biffoni
    • 5
  • Francesca Rosignolo
    • 1
  • Antonella Verrienti
    • 1
  • Marialuisa Sponziello
    • 1
  • Giuseppe Damante
    • 6
  • Diego Russo
    • 7
    Email author
  • Cosimo Durante
    • 1
  1. 1.Dipartimento di Medicina Interna e Specialità MedicheUniversità di Roma “Sapienza”RomeItaly
  2. 2.Dipartimento di Ematologia, Oncologia e Medicina MolecolareIstituto Superiore di SanitàRomeItaly
  3. 3.Dipartimento di Scienze Radiologiche, Oncologiche ed AnatomopatologicheUniversità di Roma “Sapienza”RomeItaly
  4. 4.Dipartimento di Medicina Specialistica, Diagnostica e SperimentaleUniversità di BolognaBolognaItaly
  5. 5.Dipartimento di Scienze ChirurgicheUniversità di Roma “Sapienza”RomeItaly
  6. 6.Dipartimento di Scienze Mediche e BiologicheUniversità di UdineUdineItaly
  7. 7.Dipartimento di Scienze della SaluteUniversità di Catanzaro “Magna Graecia”CatanzaroItaly

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