Reelin is a regulator of cell migration in the nervous system, and has other functions in the development of a number of non-neuronal tissues. In addition, alterations in reelin expression levels have been reported in breast, pancreatic, liver, gastric, and other cancers. Reelin is normally expressed in mammary gland stromal cells, but whether stromal reelin contributes to breast cancer progression is unknown. Herein, we used a syngeneic mouse mammary tumor transplantation model to examine the impact of host-derived reelin on breast cancer progression. We found that transplanted syngeneic tumors grew more slowly in reelin-deficient (rl Orl −/−) mice and had delayed metastatic colonization of the lungs. Immunohistochemistry of primary tumors revealed that tumors grown in rl Orl −/− animals had fewer blood vessels and increased macrophage infiltration. Gene expression studies from tumor tissues indicate that loss of host-derived reelin alters the balance of M1- and M2-associated macrophage markers, suggesting that reelin may influence the polarization of these cells. Consistent with this, rl Orl −/− M1-polarized bone marrow-derived macrophages have heightened levels of the M1-associated cytokines iNOS and IL-6. Based on these observations, we propose a novel function for the reelin protein in breast cancer progression.
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We thank Dr. Patricia Phelps, Dr. Cristina Ghiani, Dr. Catalina Abad Rabat and Dr. Diana Moughon for kindly providing reagents and for thoughtful discussion. We are thankful to Donna Crandall for assistance with figure preparation and Joseph Argus for assistance with manuscript editing.
These studies were supported by the National Institute of Child Health and Development R03 HD075840 - https://www.nichd.nih.gov/Pages/index.aspx and the California Breast Cancer Research Program 161B-0110 - http://www.cbcrp.org/to EMC. EK was supported by the Whitcome Fellowship of the UCLA Molecular Biology Institute. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Conflict of Interest
The authors declare that they have no conflict of interest.
All animal studies were conducted in accordance with the UCLA Office of Animal Research Oversight and Institutional Animal Care and Use Committee protocols.
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Metastasis of 4T1 cells in rl Orl −/− and rl Orl +/+ mice bearing similar-sized tumors. (a) Wet weight of primary tumors collected 25 days (rl Orl +/+, n = 8) and 29 days (rl Orl −/−, n = 9) after 4T1 cell injection. (b) Quantification of metastatic burden in the lungs. ns – not significant. Statistical significance determined using two-tailed, unpaired Student’s t-test. (GIF 8 kb)
Cytokine expression levels in rl Orl −/− and rl Orl +/+ BMDM treated with 4T1-conditioned medium for 24 h. Gene expression levels are relative to those in rl Orl +/+ BMDM controls. *P < 0.05, ns – not significant. Statistical significance determined using two-tailed, unpaired Student’s t-test. (GIF 6 kb)
Cytokine expression levels in rl Orl +/+ BMDM treated with IFNγ and LPS. Gene expression levels are relative to those in unstimulated rl Orl +/+ BMDM controls. (GIF 10 kb)
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Khialeeva, E., Chou, J.W., Allen, D.E. et al. Reelin Deficiency Delays Mammary Tumor Growth and Metastatic Progression. J Mammary Gland Biol Neoplasia 22, 59–69 (2017). https://doi.org/10.1007/s10911-017-9373-z