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Cancer Stem Cells
Version of Record online: 12 FEB 2013
Copyright © 2012 AlphaMed Press
Volume 31, Issue 2, pages 248–258, February 2013
How to Cite
Yang, J., Liao, D., Chen, C., Liu, Y., Chuang, T.-H., Xiang, R., Markowitz, D., Reisfeld, R. A. and Luo, Y. (2013), Tumor-Associated Macrophages Regulate Murine Breast Cancer Stem Cells Through a Novel Paracrine EGFR/Stat3/Sox-2 Signaling Pathway. STEM CELLS, 31: 248–258. doi: 10.1002/stem.1281
Author contributions: Y.L.: conception, design and manuscript writing, and final approval of manuscript; R.A.R.: manuscript writing and final approval of manuscript; J.Y. and D.L.: collection of data and data analysis; C.C., Y.L., T.-H.C., R.X., and D.M.: collection of data. J.Y. and D.L. contributed equally to this article.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS November 21, 2012.
- Issue online: 12 FEB 2013
- Version of Record online: 12 FEB 2013
- Accepted manuscript online: 21 NOV 2012 12:32AM EST
- Manuscript Accepted: 28 OCT 2012
- Manuscript Received: 30 AUG 2012
- Major State Basic Research development Program of China (973 program). Grant Number: 2013CB967202
- National Science Foundation of China (NSFC). Grant Numbers: 91029734, 81071711
- Grant Nos. Grant Numbers: 30830096, 09ZCZDSF04000
- EMD Serono Research Center. Grant Number: SFP 1645
Additional Supporting Information may be found in the online version of this article.
|sc-12-0619_sm_SupplFigure1.TIF||99K||Supplemental Figure 1. RAW macrophages possess an M2 phenotype. RAW macrophages were cultured either alone, co-cultured with 4T1 breast cancer cells or stimulated with INF- or IL-13 for 24 h, and then analyzed by flow cytometry. Macrophages were identified by CD45+/F4/80+. RAW macrophages are CD206high, IL-10high, CD86low, MHC Class II low, characteristic of M2 macrophages and TAMs.|
|sc-12-0619_sm_SupplFigure2.TIF||151K||Supplemental Figure 2. The pattern of cytokines and growth factors released in TAM (M2-macrophages). (A) The release of Th2 cytokines (IL-4, IL-10 IL-6 and TNF-α) and growth factors (EGF, VEGF and TGF-β1) were increased in TAMs derived from 4T1 tumor tissue, reversely, Th1 cytokines (IL-2, IL-7, IL12a, IL-12b and INF-r) were released largly in MI macrophages derived from spleen of normal mice. (B-C) The expression of cytokines (IL-4, IL-10 IL-6 and TNF-α) and growth factors (EGF, VEGF and TGF-β1) were up-regulated in RAW cells after co-cultured with 4T1 tumor cells or 4T1 tumor conditional medium. (D) The expression of growth factors (TNF-α, EGF, VEGF and TGF-β1) were up-regulated in 4T1 cells after co-cultured with RAW cells.|
|sc-12-0619_sm_SupplFigure3.TIF||113K||Supplemental Figure 3. 4T1 and 4T07 breast cancer cells contain a side population (SP) with cancer stem cell-like properties. (A) 4T07 or 4T1 breast cancer cells were first stained with Hoechst 3334 dye, and then reacted with either Sca-1 or ABCG2 antibodies labeled with PE to determine the SP and its expression of Sca-1 and ABCG2. (B) SP of 4T07 cells was sorted by Flow Cytometry after being stained with Hoechst 3334 dye, and a group of Balb/c mice were implanted in the cleared fat pad with either the SP or Non-SP fraction of 4T07 breast cancer cells at 7×103 per mouse. (C) Tumor volume was measured every 3-5 days. (D) All mice were sacrificed on day 25 after tumor cell implantation and analyzed for the metastasis score on lungs|
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