Tumor initiating cells: Development and critical characterization of a model derived from the A431 carcinoma cell line forming spheres in suspension

Ileana Bortolomai, Silvana Canevari, Ileana Facetti, Loris De Cecco, Giancarlo Castellano, Alberto Zacchetti, Malcolm R. Alison, Silvia Miotti

Research output: Contribution to journalArticle

Abstract

To investigate the tumor fraction with cancer stem/tumor initiating cell (CSC/TIC) characteristics, we tested the human cervical carcinoma cell lines A431, Caski and SiHa, by growth as non-adherent spheres in specific media and aldehyde dehydrogenase (ALDH) enzymatic activity. A good correlation between the two parameters was observed and the highest levels were observed in A431 cell line that was selected for characterization of the CSC/TIC fraction. A431 parental cells already displayed characteristics common to CSC/TIC, such as sphere forming efficiency, adherent holoclone formation and high ALDH activity. Non-adherent spheres maintained or increased these properties, and, in particular, ALDH-positive fraction increased from 46 to 65% and a transient induction of stem cell markers such as Nanog, Nestin and Oct4 was observed. Furthermore, a significant increase of paraclone forming cells was observed, suggesting that differentiation took place inside sphere cell populations. As compared to parental cells, spheres were characterized by: (1) a ten-fold higher verapamil-sensitive side population fraction; (2) the appearance of a podoplanin-positive subpopulation characterized by a small cell size; (3) the ability to propagate tumors in nude mice at a lower cell dose. The global gene expression analysis demonstrated a strong and reversible modulation of 'sphere' phenotype in comparison to parental and sphere cells re-induced to adherent conditions. All together our results indicated that the growth of A431 cells as a non-adherent sphere was not sufficient by itself to define a stem-like population, but it was essential for the emergence of a small population of tumor cells with CSC properties.

Original languageEnglish
Pages (from-to)1194-1206
Number of pages13
JournalCell Cycle
Volume9
Issue number6
Publication statusPublished - Mar 15 2010

Fingerprint

Neoplastic Stem Cells
Suspensions
Carcinoma
Cell Line
Aldehyde Dehydrogenase
Neoplasms
Population
Nestin
Vulnerable Populations
Growth
Verapamil
Cell Size
Nude Mice
Stem Cells
Phenotype
Gene Expression

Keywords

  • ALDH
  • Cancer stem cells
  • Gene profiling
  • Holoclones
  • Podoplanin
  • Side population
  • Sphere formation

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Developmental Biology

Cite this

Bortolomai, I., Canevari, S., Facetti, I., De Cecco, L., Castellano, G., Zacchetti, A., ... Miotti, S. (2010). Tumor initiating cells: Development and critical characterization of a model derived from the A431 carcinoma cell line forming spheres in suspension. Cell Cycle, 9(6), 1194-1206.

Tumor initiating cells : Development and critical characterization of a model derived from the A431 carcinoma cell line forming spheres in suspension. / Bortolomai, Ileana; Canevari, Silvana; Facetti, Ileana; De Cecco, Loris; Castellano, Giancarlo; Zacchetti, Alberto; Alison, Malcolm R.; Miotti, Silvia.

In: Cell Cycle, Vol. 9, No. 6, 15.03.2010, p. 1194-1206.

Research output: Contribution to journalArticle

Bortolomai, I, Canevari, S, Facetti, I, De Cecco, L, Castellano, G, Zacchetti, A, Alison, MR & Miotti, S 2010, 'Tumor initiating cells: Development and critical characterization of a model derived from the A431 carcinoma cell line forming spheres in suspension', Cell Cycle, vol. 9, no. 6, pp. 1194-1206.
Bortolomai, Ileana ; Canevari, Silvana ; Facetti, Ileana ; De Cecco, Loris ; Castellano, Giancarlo ; Zacchetti, Alberto ; Alison, Malcolm R. ; Miotti, Silvia. / Tumor initiating cells : Development and critical characterization of a model derived from the A431 carcinoma cell line forming spheres in suspension. In: Cell Cycle. 2010 ; Vol. 9, No. 6. pp. 1194-1206.
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