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. 2014 May 15;7(6):2934-41.
eCollection 2014.

MAGE-A3 is highly expressed in a cancer stem cell-like side population of bladder cancer cells

Bo Yin  1 , Yu Zeng  2 , Gang Liu  1 , Xiaotian Wang  1 , Peng Wang  1 , Yongsheng Song  1
Affiliations

MAGE-A3 is highly expressed in a cancer stem cell-like side population of bladder cancer cells

Bo Yin et al. Int J Clin Exp Pathol. .

Abstract

Cancer stem cells (CSCs), which have the abilities of tumor-initiating, self-renewal and differentiation, are thought to cause post-therapeutic recurrence and the progression of cancer. However, CSCs are commonly resistant to current cancer therapies including chemotherapy and radiotherapy. In this study, we isolated cancer stem celllike side population (SP) cells from human bladder cancer cell line SW780 by a flow cytometry-based SP technique. SP cells were only about 3.6% of SW780 cells and showed higher expression of ATP-binding cassette sub-family G member 2 (ABCG2) and CD133. In vitro assay of tumor sphere growth as well as in vivo assay of xenograft transplantation confirmed the higher tumorigenicity of isolated SP cells. These data indicated that SP cells were enriched with CSCs of bladder cancer. Furthermore, we determined the expression of melanoma antigen family A, 3 (MAGEA3), one of the most studied cancer testis (CT) antigens, in these SP and main population (MP) cells derived from SW780 cells. SW780 SP cells representing CSCs of bladder cancer showed an up-regulated expression of MAGE-A3 and a positive coexpression of MAGE-A3 and CD133, indicating that MAGE-A3 was a novel CT antigen preferentially expressed in the CSCs of bladder cancer. In summary, our findings confirmed the existence of cancer stem cell-like SP cells in bladder cancer cells, and further indicated that MAGE-A3 is a novel CSC antigen and therefore may serve as an immunotherapeutic target for CSCs of bladder cancer.

Keywords: 3 (MAGE-A3); Melanoma antigen family A; bladder cancer; cancer stem cells; cancertestis; immunotherapy; side population.

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Figures

Figure 1
Figure 1
Isolation of SP cells from SW780 cell line by Hoechst 33342 SP analysis in the absence (A) or presence (B) of verapamil.
Figure 2
Figure 2
Expression of ABCG2 (A) and CD133 (B) detected by RT-PCR.
Figure 3
Figure 3
In vitro growth characteristics of SW780 SP cells. SP cells grew into sphere-like clusters/tumor spheres, as illustrated by phase contrast microscopy (20 ×).
Figure 4
Figure 4
Higher in vivo tumorigenicity of SW780 SP cells. Data are means ± SD.
Figure 5
Figure 5
Expression of MAGE-A3 in cancer stem cell-like SP cells. Expression of MAGE-A3 was detected by RT-PCR (A) and Western blot (B); Double immunofluorescence analysis (C) of coexpression of MAGE-A3 (red) and CD133 (green) was performed in SW780 SP cells. DAPI was used to stain nuclei (200 × magnification).
See this image and copyright information in PMC

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