Effects of cytokines derived from cancer-associated fibroblasts on androgen synthetic enzymes in estrogen receptor-negative breast carcinoma
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The tumor microenvironment plays pivotal roles in promotion of many malignancies. Cancer-associated fibroblasts (CAFs) have been well-known to promote proliferation, angiogenesis, and metastasis but mechanistic understanding of tumor–stroma interactions is not yet complete. Recently, estrogen synthetic enzymes were reported to be upregulated by co-culture with stromal cells in ER positive breast carcinoma (BC) but effects of co-culture on androgen metabolism have not been extensively examined. Therefore, we evaluated roles of CAFs on androgen metabolism in ER-negative AR-positive BC through co-culture with CAFs.
Concentrations of steroid hormone in supernatant of co-culture of MDA-MB-453 and primary CAFs were measured using GC–MS. Cytokines derived from CAFs were determined using Cytokine Array. Expressions of androgen synthetic enzymes were confirmed using RT-PCR and Western blotting. Correlations between CAFs and androgen synthetic enzymes were analyzed using triple-negative BC (TNBC) patient tissues by immunohistochemistry.
CAFs were demonstrated to increase expressions and activities of 17βHSD2, 17βHSD5, and 5α-Reductase1. IL-6 and HGF that were selected as potential paracrine mediators using cytokine array induced 17βHSD2, 17βHSD5, and 5α-Reductase1 expression. Underlying mechanisms of IL-6 paracrine regulation of 17βHSD2 and 17βHSD5 could be partially dependent on phosphorylated STAT3, while phosphorylated ERK could be involved in HGF-mediated 5α-Reductase1 induction. α-SMA status was also demonstrated to be significantly correlated with 17βHSD2 and 17βHSD5 status in TNBC tissues, especially AR-positive cases.
Results of our present study suggest that both IL-6 and HGF derived from CAFs could contribute to the intratumoral androgen metabolism in ER-negative BC patients.
KeywordsBreast cancer Triple-negative breast cancer (TNBC) Microenvironment Cancer-associated fibroblasts (CAFs) Androgen
Human epidermal growth factor receptor
Gas chromatography–mass spectrometry
Reverse transcription polymerase chain reaction
Triple-negative breast carcinoma
17β hydroxysteroid dehydrogenase type
- IL-6 (-8)
Hepatocyte growth factor,
Signal transducer and activator of transcription 3
Extracellular signal-regulated kinase
Invasive ductal carcinoma
Α-smooth muscle actin
Fetal bovine serum
Plasminogen activator inhibitor-1
Monocyte chemotactic protein-1
Small interfering RNA
Fatty acid synthase
Research for this article was supported in part by JSPS KAKENHI Grant Number 15K18396. We would like to acknowledge all the members of their laboratories, whose informal input was extremely valuable. We would also like to acknowledge the support and assistance of the members of the Department of Pathology, Tohoku University School of Medicine.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This study was approved by Ethics Committee at Tohoku University School of Medicine. Informed consent was obtained from all patients.
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