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 language | English |
|---|---|
| Pages (from-to) | 1194-1206 |
| Number of pages | 13 |
| Journal | Cell Cycle |
| Volume | 9 |
| Issue number | 6 |
| Publication status | Published - Mar 15 2010 |
Fingerprint
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
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 journal › Article
}
TY - JOUR
T1 - Tumor initiating cells
T2 - Development and critical characterization of a model derived from the A431 carcinoma cell line forming spheres in suspension
AU - Bortolomai, Ileana
AU - Canevari, Silvana
AU - Facetti, Ileana
AU - De Cecco, Loris
AU - Castellano, Giancarlo
AU - Zacchetti, Alberto
AU - Alison, Malcolm R.
AU - Miotti, Silvia
PY - 2010/3/15
Y1 - 2010/3/15
N2 - 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.
AB - 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.
KW - ALDH
KW - Cancer stem cells
KW - Gene profiling
KW - Holoclones
KW - Podoplanin
KW - Side population
KW - Sphere formation
UR - http://www.scopus.com/inward/record.url?scp=77953521844&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77953521844&partnerID=8YFLogxK
M3 - Article
C2 - 20237414
AN - SCOPUS:77953521844
VL - 9
SP - 1194
EP - 1206
JO - Cell Cycle
JF - Cell Cycle
SN - 1538-4101
IS - 6
ER -