Distinct nuclear receptor expression in stroma adjacent to breast tumors
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The interaction between breast tumor epithelial and stromal cells is vital for initial and recurrent tumor growth. While breast cancer-associated stromal cells provide a favorable environment for proliferation and metastasis, the molecular mechanisms contributing to this process are not fully understood. Nuclear receptors (NRs) are intracellular transcription factors that directly regulate gene expression. Little is known about the status of NRs in cancer-associated stroma. Nuclear Receptor Low-Density Taqman Arrays were used to compare the gene expression profiles of all 48 NR family members in a collection of primary cultured cancer-associated fibroblasts (CAFs) obtained from estrogen receptor (ER)α positive breast cancers (n = 9) and normal breast adipose fibroblasts (NAFs) (n = 7). Thirty-three of 48 NRs were expressed in both the groups, while 11 NRs were not detected in either. Three NRs (dosage-sensitive sex reversal, adrenal hypoplasia critical region, on chromosome X, gene 1 (DAX-1); estrogen-related receptor beta (ERR-β); and RAR-related orphan receptor beta (ROR-β)) were only detected in NAFs, while one NR (liver receptor homolog-1 (LRH-1)) was unique to CAFs. Of the NRs co-expressed, four were significantly down-regulated in CAFs compared with NAFs (RAR-related orphan receptor-α (ROR-α); Thyroid hormone receptor-β (TR-β); vitamin D receptor (VDR); and peroxisome proliferator-activated receptor-γ (PPAR-γ)). Quantitative immunohistochemistry for LRH-1, TR-β, and PPAR-γ proteins in stromal fibroblasts from an independent panel of breast cancers (ER-positive (n = 15), ER-negative (n = 15), normal (n = 14)) positively correlated with mRNA expression profiles. The differentially expressed NRs identified in tumor stroma are key mediators in aromatase regulation and subsequent estrogen production. Our findings reveal a distinct pattern of NR expression that therefore fits with a sustained and increased local estrogen microenvironment in ER-positive tumors. NRs in CAFs may provide a new avenue for the development of intratumoral-targeted therapies in breast cancer.
KeywordsNuclear receptors Breast cancer Stroma Aromatase Estrogen Tumor microenvironment
This study was supported by the National Health and Medical Research Council of Australia through fellowships to CDC (#338518), JEV (Australia Fellow), and GJL (637307), ERS, PJF, and CLC; and NHMRC IRIISS; GEOM is the University of Queensland Vice Chancellors research fellowship; the Victorian State Government through Victoria Cancer Agency funding of the Victoria Breast Cancer Research Consortium and the Victorian Government’s Operational Infrastructure Support Program; the United States Department of Defense fellowship to ALC; a Collaborative Program from the National Breast Cancer Foundation Australia. Breast cancer and normal tissues were provided by Australian Breast Cancer Tissue Bank (ABCTB), which is supported by the National Health and Medical Research Council of Australia, the Cancer Institute NSW, and the National Breast Cancer Foundation, or by the Victorian Cancer BioBank Australia, which is supported by the Victorian Government. ABCTB tissues and samples were made available to researchers on a nonexclusive basis. Normal breast biopsies were obtained from the Susan G. Komen for the Cure Tissue Bank at the IU Simon Cancer Center. The authors thank contributors to the Susan G. Komen for the Cure Tissue Bank, including Indiana University that collected the samples used in this study, as well as parents and families whose participation and help made this study possible. The authors would like to thank Drs Elgene Lim and François Vaillant for their assistance in stromal cell collection, and Silke Kantimm for assistance with immunohistochemistry. PHI Data Audit #13-08.
Conflict of interest
The authors declare no conflicts of interest.
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