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Identification and characterization of cancer stem cells in human head and neck squamous cell carcinoma.

Han J, Fujisawa T, Husain SR, Puri RK - BMC Cancer (2014)

Bottom Line: Our results show that CD24+/CD44+ cells possessed stemness characteristics of self-renewal and differentiation.In addition, the CD24+/CD44+ cells were more chemo-resistant to gemcitabine and cisplatin compared to CD24-/CD44+ cells.In vivo, CD24+/CD44+ cells showed a tendency to generate larger tumors in nude mice compared to CD24-/CD44+ cell population.

View Article: PubMed Central - HTML - PubMed

Affiliation: Tumor Vaccines and Biotechnology Branch, Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, NIH Bldg 29B, Rm 2NN20, 29 Lincoln Dr,, Bethesda, MD, 20892, USA. raj.puri@fda.hhs.gov.

ABSTRACT

Background: Current evidence suggests that initiation, growth, and invasion of cancer are driven by a small population of cancer stem cells (CSC). Previous studies have identified CD44+ cells as cancer stem cells in head and neck squamous cell carcinoma (HNSCC). However, CD44 is widely expressed in most cells in HNSCC tumor samples and several cell lines tested. We previously identified a small population of CD24+/CD44+ cells in HNSCC. In this study, we examined whether this population of cells may represent CSC in HNSCC.

Methods: CD24+/CD44+ cells from HNSCC cell lines were sorted by flow cytometry, and their phenotype was confirmed by qRT-PCR. Their self-renewal and differentiation properties, clonogenicity in collagen gels, and response to anticancer drugs were tested in vitro. The tumorigenicity potential of CD24+/CD44+ cells was tested in athymic nude mice in vivo.

Results: Our results show that CD24+/CD44+ cells possessed stemness characteristics of self-renewal and differentiation. CD24+/CD44+ cells showed higher cell invasion in vitro and made higher number of colonies in collagen gels compared to CD24-/CD44+ HNSCC cells. In addition, the CD24+/CD44+ cells were more chemo-resistant to gemcitabine and cisplatin compared to CD24-/CD44+ cells. In vivo, CD24+/CD44+ cells showed a tendency to generate larger tumors in nude mice compared to CD24-/CD44+ cell population.

Conclusion: Our study clearly demonstrates that a distinct small population of CD24+/CD44+ cells is present in HNSCC that shows stem cell-like properties. This distinct small population of cells should be further characterized and may provide an opportunity to target HNSCC CSC for therapy.

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Representative flow cytometry plots analyzing expression of CD29, CD24, CD44, and CD73 in A253 HNSCC cells. (A) Co-expression of CD29 with CD44 and CD24. (B) Co-expression of CD73 with CD44 and CD24.
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Figure 8: Representative flow cytometry plots analyzing expression of CD29, CD24, CD44, and CD73 in A253 HNSCC cells. (A) Co-expression of CD29 with CD44 and CD24. (B) Co-expression of CD73 with CD44 and CD24.

Mentions: To investigate whether other putative stem cell markers were expressed in HNSCC cells, the mesenchymal stem cell markers, CD29 (β1-integrin), CD73 (5′-nucleotidase), CD90 (Thy-1), and CD105 (Endogin) were selected and analyzed by flow cytometry. CD29 expression showed the strongest correlation with the CD44 expression. Almost all cells (99.6%) were CD29 and CD44 double-positive. Only ~6% of the cells were CD29+ and CD24+, the same percentage found for CD24+/CD44+ cells (Figure 8A). CD73 also showed a strong correlation with CD44 expression. Approximately 92% of the cells were CD73 and CD44 double-positive, while only ~6% of the cells were CD73+/CD24+, similar to CD24+/CD44+ cells (Figure 8B). On the other hand, neither CD90 nor CD105 showed any correlation with either CD24 or CD44 expression (data not shown).


Identification and characterization of cancer stem cells in human head and neck squamous cell carcinoma.

Han J, Fujisawa T, Husain SR, Puri RK - BMC Cancer (2014)

Representative flow cytometry plots analyzing expression of CD29, CD24, CD44, and CD73 in A253 HNSCC cells. (A) Co-expression of CD29 with CD44 and CD24. (B) Co-expression of CD73 with CD44 and CD24.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4008349&req=5

Figure 8: Representative flow cytometry plots analyzing expression of CD29, CD24, CD44, and CD73 in A253 HNSCC cells. (A) Co-expression of CD29 with CD44 and CD24. (B) Co-expression of CD73 with CD44 and CD24.
Mentions: To investigate whether other putative stem cell markers were expressed in HNSCC cells, the mesenchymal stem cell markers, CD29 (β1-integrin), CD73 (5′-nucleotidase), CD90 (Thy-1), and CD105 (Endogin) were selected and analyzed by flow cytometry. CD29 expression showed the strongest correlation with the CD44 expression. Almost all cells (99.6%) were CD29 and CD44 double-positive. Only ~6% of the cells were CD29+ and CD24+, the same percentage found for CD24+/CD44+ cells (Figure 8A). CD73 also showed a strong correlation with CD44 expression. Approximately 92% of the cells were CD73 and CD44 double-positive, while only ~6% of the cells were CD73+/CD24+, similar to CD24+/CD44+ cells (Figure 8B). On the other hand, neither CD90 nor CD105 showed any correlation with either CD24 or CD44 expression (data not shown).

Bottom Line: Our results show that CD24+/CD44+ cells possessed stemness characteristics of self-renewal and differentiation.In addition, the CD24+/CD44+ cells were more chemo-resistant to gemcitabine and cisplatin compared to CD24-/CD44+ cells.In vivo, CD24+/CD44+ cells showed a tendency to generate larger tumors in nude mice compared to CD24-/CD44+ cell population.

View Article: PubMed Central - HTML - PubMed

Affiliation: Tumor Vaccines and Biotechnology Branch, Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, NIH Bldg 29B, Rm 2NN20, 29 Lincoln Dr,, Bethesda, MD, 20892, USA. raj.puri@fda.hhs.gov.

ABSTRACT

Background: Current evidence suggests that initiation, growth, and invasion of cancer are driven by a small population of cancer stem cells (CSC). Previous studies have identified CD44+ cells as cancer stem cells in head and neck squamous cell carcinoma (HNSCC). However, CD44 is widely expressed in most cells in HNSCC tumor samples and several cell lines tested. We previously identified a small population of CD24+/CD44+ cells in HNSCC. In this study, we examined whether this population of cells may represent CSC in HNSCC.

Methods: CD24+/CD44+ cells from HNSCC cell lines were sorted by flow cytometry, and their phenotype was confirmed by qRT-PCR. Their self-renewal and differentiation properties, clonogenicity in collagen gels, and response to anticancer drugs were tested in vitro. The tumorigenicity potential of CD24+/CD44+ cells was tested in athymic nude mice in vivo.

Results: Our results show that CD24+/CD44+ cells possessed stemness characteristics of self-renewal and differentiation. CD24+/CD44+ cells showed higher cell invasion in vitro and made higher number of colonies in collagen gels compared to CD24-/CD44+ HNSCC cells. In addition, the CD24+/CD44+ cells were more chemo-resistant to gemcitabine and cisplatin compared to CD24-/CD44+ cells. In vivo, CD24+/CD44+ cells showed a tendency to generate larger tumors in nude mice compared to CD24-/CD44+ cell population.

Conclusion: Our study clearly demonstrates that a distinct small population of CD24+/CD44+ cells is present in HNSCC that shows stem cell-like properties. This distinct small population of cells should be further characterized and may provide an opportunity to target HNSCC CSC for therapy.

Show MeSH
Related in: MedlinePlus