Isolation of head and neck squamous carcinoma cancer stem-like cells in a syngeneic mouse model and analysis of hypoxia effect

Duarte,Sónia; Loubat,Agnès; Momier,David; Topi,Majlinda; Faneca,Henrique; Pedroso de Lima,Maria   C.; Carle,Georges  F.; Pierrefite-Carle,Valérie

doi:10.3892/or.2012.1904

Isolation of head and neck squamous carcinoma cancer stem-like cells in a syngeneic mouse model and analysis of hypoxia effect

Authors:
- Sónia Duarte
- Agnès Loubat
- David Momier
- Majlinda Topi
- Henrique Faneca
- Maria C. Pedroso de Lima
- Georges F. Carle
- Valérie Pierrefite-Carle
View Affiliations

Affiliations: CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal, IFR50, School of Medicine, University of Nice Sophia Antipolis, Nice, France, GéPITOs UMR 6235 CNRS, School of Medicine, University of Nice Sophia Antipolis, Nice, France
Published online on: July 6, 2012 https://doi.org/10.3892/or.2012.1904
Pages: 1057-1062

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The incidence of oral tumors is increasing around the world and despite recent advances in early detection and diagnosis, current treatments are still unsatisfactory. Recent data suggest that tumor persistence and recurrence could be due to the presence of a rare cell population called cancer stem cells (CSCs), which are generally spared by traditional treatments. Therefore, identification and characterization of CSCs are extremely important to develop novel and effective treatment strategies for cancer. The aim of this study was to identify and isolate CSCs in an established murine head and neck squamous cell carcinoma (HNSCC) cell line and to investigate the influence of hypoxic conditions on the isolated cell population. Using the expression of the aldehyde dehydrogenase 1 (ALDH1) enzymatic activity, which is now recognized as a CSC marker in various tumors, we isolated a cell population expressing high levels of ALDH1 (ALDH1high) representing 1±0.6% in the murine SCC-VII cell line. These cells were injected subcutaneously in syngeneic animals to evaluate their tumorigenic properties. For the lowest injected cell dose (250 injected cells), tumor occurrence and median tumor size were higher in ALDH1high injected mice than in ALDH1low injected mice. Following an in vivo passage and culture in serum-free medium, the percentage of ALDH1high cells increased by 3‑fold in SCC-VII CSCs (oral spheres) compared to the SCC-VII cell line. This percentage was further increased when oral spheres were cultured under hypoxic conditions. In conclusion, this study reports for the first time the isolation of HNSCC CSCs in a syngeneic mouse model and the use of hypoxia as a method to further enrich the ALDH1high cell population.

View Figures

View References

1	Argiris A, Karamouzis MV, Raben D and Ferris RL: Head and neck cancer. Lancet. 371:1695–1709. 2008. View Article : Google Scholar
2	Moore SR, Johnson NW, Pierce AM and Wilson DF: The epidemiology of mouth cancer: a review of global incidence. Oral Dis. 6:65–74. 2000. View Article : Google Scholar : PubMed/NCBI
3	Forastiere A, Koch W, Trotti A and Sidransky D: Head and neck cancer. N Engl J Med. 345:1890–1900. 2001. View Article : Google Scholar
4	Forastiere AA, Trotti A, Pfister DG and Grandis JR: Head and neck cancer: recent advances and new standards of care. J Clin Oncol. 24:2603–2605. 2006. View Article : Google Scholar : PubMed/NCBI
5	Visvader JE and Lindeman GJ: Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer. 8:755–768. 2008. View Article : Google Scholar : PubMed/NCBI
6	Tabor MH, Clay MR, Owen JH, et al: Head and neck cancer stem cells: the side population. Laryngoscope. 121:527–533. 2011. View Article : Google Scholar : PubMed/NCBI
7	Li HZ, Yi TB and Wu ZY: Suspension culture combined with chemotherapeutic agents for sorting of breast cancer stem cells. BMC Cancer. 8:1352008. View Article : Google Scholar : PubMed/NCBI
8	Prince ME, Sivanandan R, Kaczorowski A, et al: Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci USA. 104:973–978. 2007. View Article : Google Scholar : PubMed/NCBI
9	Lim YC, Oh SY, Cha YY, Kim SH, Jin X and Kim H: Cancer stem cell traits in squamospheres derived from primary head and neck squamous cell carcinomas. Oral Oncol. 47:83–91. 2011. View Article : Google Scholar : PubMed/NCBI
10	Chen H, Zhou L, Dou T, Wan G, Tang H and Tian J: BMI1’s maintenance of the proliferative capacity of laryngeal cancer stem cells. Head Neck. 33:1115–1125. 2011.
11	Chen YC, Chen YW, Hsu HS, et al: Aldehyde dehydrogenase 1 is a putative marker for cancer stem cells in head and neck squamous cancer. Biochem Biophys Res Commun. 385:307–313. 2009. View Article : Google Scholar : PubMed/NCBI
12	Clay MR, Tabor M, Owen JH, et al: Single-marker identification of head and neck squamous cell carcinoma cancer stem cells with aldehyde dehydrogenase. Head Neck. 32:1195–1201. 2010. View Article : Google Scholar : PubMed/NCBI
13	Ginestier C, Hur MH, Charafe-Jauffret E, et al: ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell. 1:555–567. 2007. View Article : Google Scholar : PubMed/NCBI
14	Quintana E, Shackleton M, Sabel MS, Fullen DR, Johnson TM and Morrison SJ: Efficient tumour formation by single human melanoma cells. Nature. 456:593–598. 2008. View Article : Google Scholar : PubMed/NCBI
15	Auerbach R, Morrissey LW and Sidky YA: Regional differences in the incidence and growth of mouse tumors following intradermal or subcutaneous inoculation. Cancer Res. 38:1739–1744. 1978.PubMed/NCBI
16	Chen ZG: The cancer stem cell concept in progression of head and neck cancer. J Oncol. 2009:8940642009.PubMed/NCBI
17	Frank NY, Schatton T and Frank MH: The therapeutic promise of the cancer stem cell concept. J Clin Invest. 120:41–50. 2010. View Article : Google Scholar : PubMed/NCBI
18	Sayed SI, Dwivedi RC, Katna R, et al: Implications of understanding cancer stem cell (CSC) biology in head and neck squamous cell cancer. Oral Oncol. 47:237–243. 2011. View Article : Google Scholar : PubMed/NCBI
19	Dontu G, Abdallah WM, Foley JM, et al: In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev. 17:1253–1270. 2003. View Article : Google Scholar : PubMed/NCBI
20	Ricci-Vitiani L, Lombardi DG, Pilozzi E, et al: Identification and expansion of human colon-cancer-initiating cells. Nature. 445:111–115. 2007. View Article : Google Scholar : PubMed/NCBI
21	Cabarcas SM, Mathews LA and Farrar WL: The cancer stem cell niche - there goes the neighborhood? Int J Cancer. 129:2315–2327. 2011. View Article : Google Scholar : PubMed/NCBI
22	Das B, Tsuchida R, Malkin D, Koren G, Baruchel S and Yeger H: Hypoxia enhances tumor stemness by increasing the invasive and tumorigenic side population fraction. Stem Cells. 26:1818–1830. 2008. View Article : Google Scholar : PubMed/NCBI
23	Yeung TM, Gandhi SC and Bodmer WF: Hypoxia and lineage specification of cell line-derived colorectal cancer stem cells. Proc Natl Acad Sci USA. 108:4382–4387. 2011. View Article : Google Scholar : PubMed/NCBI