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. 2013 Jun 24;13(1):62.
doi: 10.1186/1475-2867-13-62.

Increased invasion and tumorigenicity capacity of CD44+/CD24- breast cancer MCF7 cells in vitro and in nude mice

Wenxing Yan  1 , Yubing Chen, Yueliang Yao, Hongmei Zhang, Tiejun Wang
Affiliations

Increased invasion and tumorigenicity capacity of CD44+/CD24- breast cancer MCF7 cells in vitro and in nude mice

Wenxing Yan et al. Cancer Cell Int. .

Abstract

Background: Identification of cancer stem cells (CSCs) and their behaviors will provide insightful information for the future control of human cancers. This study investigated CD44 and CD24 cell surface markers as breast cancer CSC markers in vitro and in vivo.

Methods: Flow cytometry with CD44 and CD24 markers was used to sort breast cancer MCF7 cells for scanning electron microscopy (SEM), tumor cell invasion assay, and nude mouse xenograft assay.

Results: Flow cytometry assay using CD44 and CD24 markers sorted MCF7 cells into four subsets, i.e., CD44+/CD24-/low, CD44-/CD24+, CD44+/CD24+, and CD44-/CD24-. The SEM data showed that there were many protrusions on the surface of CD44+/CD24-/low cells. CD44+/CD24-/low cells had many microvilli and pseudopodia. The CD44+/CD24-/low cells had a higher migration and invasion abilities than that of the other three subsets of the cells. The in vivo tumor formation assay revealed that CD44+/CD24- cells had the highest tumorigenic capacity compared to the other three subsets.

Conclusion: CD44 and CD24 could be useful markers for identification of breast CSCs because CD44+/CD24-/low cells had unique surface ultrastructures and the highest tumorigenicity and invasive abilities.

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Figures

Figure 1
Figure 1
Flow cytometry sorting of MCF7 cells using CD44 and CD24 markers. A, MCF7 cells were analyzed by fluorescence-activated cell sorting (FACS) using anti-CD44 and anti-CD24 antibodies. B, Quantification of the four subsets of MCF7 cells. C, Confirmation of the purity of the CD44+/CD24-/low cell type using flow cytometry.
Figure 2
Figure 2
Cell surface ultrastructure of the four different subsets of MCF7 cells. A, CD44+/CD24-/low; B, CD44+/CD24+; C, CD44-/CD24+; and D, CD44-/CD24-.
Figure 3
Figure 3
Invasion assay of these four subsets of MCF7 cells. A, Cell number quantification of invasion. B, Confocal images of these four subsets of MCF7 cells (Green: CD44+/CD24-/low; Red: CD44-/CD24+; Yellow: CD44+/CD24+; and Blue: CD44-/CD24-). C, Quantification of tumor cell migration distance. D, Images of cell migration distance.
Figure 4
Figure 4
In vivo tumor xenograft assay of these four subsets MCF7 cells. A, CD44+/CD24-/low; B, CD44+/CD24+; C, CD44-/CD24+; and D, CD44-/CD24-.
See this image and copyright information in PMC

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