Effect of fibroblasts on breast cancer cell mammosphere formation and regulation of stem cell-related gene expression

  • Authors:
    • Fengchun Zhang
    • Caili Song
    • Yue Ma
    • Lei Tang
    • Yingchun Xu
    • Hongxia Wang
  • View Affiliations

  • Published online on: May 13, 2011     https://doi.org/10.3892/ijmm.2011.700
  • Pages: 365-371
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Abstract

The purpose of this study was to investigate the regulatory effects of breast cancer fibroblasts (BCFs) vs. normal mammary fibroblasts (NMFs) on mammosphere formation and stem cell-related gene expression in breast cancer cells. Breast cancer cells (MCF-7) were cultured in suspension to generate primary and secondary mammospheres. The proportion of CD44+/CD24low/- cells was assessed by flow cytometry (FCM), and Wnt1, Notch1, β-catenin, CXCR4, SOX2 and ALDH3A1 gene expression was detected by quantitative real-time PCR. The fibroblasts from either breast cancer tissue or normal mammary tissue were purified from tissue specimens and co-cultured with breast cancer cells. The mammosphere formation efficacy was approximately 180/10,000 MCF-7 cells. FCM analysis showed that, compared to the 2.1% positive expression in the MCF-7 monolayer culture cells, the expression of CD44+/CD24low/- in MCF-7 mammosphere cells was significantly elevated to 10.4% (P<0.01). The proportion of the CD44+/CD24low/- subpopulation of the cells in mammospheres was nearly 5-fold higher than that of general MCF-7 cells. Compared with MCF-7 monolayer culture cells, mammosphere cells showed significantly (P<0.01) enhanced expression of Wnt1 [fold-change (FC), 2.25], Notch1 (FC, 2.45), β-catenin (FC, 1.72), CXCR4 (FC, 4.68), SOX2 (FC, 4.25) and ALDH3A1 (FC, 5.38). When BCFs were co-cultured with MCF-7 cells under mammosphere culture conditions, the length of time of mammosphere formation decreased, the volume of the mammo-spheres increased and the mammosphere-forming efficiency (MFE) was higher than that of NMFs and the control group. Both the BCF and NMF groups showed enhanced gene expression for the following genes: Wnt1 (FC, 3.18 and 1.27, respectively), β-catenin (FC, 1.75 and 1.22, respectively), Notch1 (FC, 2.09 and 1.31, respectively), CXCR4 (FC, 2.77 and 1.33, respectively), SOX2 (FC, 2.77 and 1.80, respectively) and ALDH3A1 (FC, 5.23 and 1.85, respectively). Cancer fibroblast cells can promote the MFE and up-regulate stem cell-related gene expression in breast cancer cells.

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September 2011
Volume 28 Issue 3

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Zhang F, Song C, Ma Y, Tang L, Xu Y and Wang H: Effect of fibroblasts on breast cancer cell mammosphere formation and regulation of stem cell-related gene expression. Int J Mol Med 28: 365-371, 2011
APA
Zhang, F., Song, C., Ma, Y., Tang, L., Xu, Y., & Wang, H. (2011). Effect of fibroblasts on breast cancer cell mammosphere formation and regulation of stem cell-related gene expression. International Journal of Molecular Medicine, 28, 365-371. https://doi.org/10.3892/ijmm.2011.700
MLA
Zhang, F., Song, C., Ma, Y., Tang, L., Xu, Y., Wang, H."Effect of fibroblasts on breast cancer cell mammosphere formation and regulation of stem cell-related gene expression". International Journal of Molecular Medicine 28.3 (2011): 365-371.
Chicago
Zhang, F., Song, C., Ma, Y., Tang, L., Xu, Y., Wang, H."Effect of fibroblasts on breast cancer cell mammosphere formation and regulation of stem cell-related gene expression". International Journal of Molecular Medicine 28, no. 3 (2011): 365-371. https://doi.org/10.3892/ijmm.2011.700