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.
- 1.Orimo A, Gupta P, Sgroi D, Arenzana-Seisdedos F, Delaunay T, Naeem R, Carey V, Richardson A, Weinberg R (2005) Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion. Cell 121(3):335–348. doi: 10.1016/j.cell.2005.02.034X CrossRefPubMedGoogle Scholar
- 3.Dagouassat M, Suffee N, Hlawaty H, Haddad O, Charni F, Laguillier C, Vassy R, Martin L, Schischmanoff P, Gattegno L, Oudar O, Sutton A, Charnaux N (2010) Monocyte chemoattractant protein-1 (MCP-1)/CCL2 secreted by hepatic myofibroblasts promotes migration and invasion of human hepatoma cells. Int J Cancer 126(5):1095–1108. doi: 10.1002/ijc.24800 PubMedGoogle Scholar
- 5.Tsuyada A, Chow A, Wu J, Somlo G, Chu P, Loera S, Luu T, Li A, Wu X, Ye W, Chen S, Zhou W, Yu Y, Wang Y, Ren X, Li H, Scherle P, Kuroki Y, Wang S (2012) CCL2 mediates cross-talk between cancer cells and stromal fibroblasts that regulates breast cancer stem cells. Cancer Res 72(11):2768–2779. doi: 10.1158/0008-5472.CAN-11-3567 CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Wang X, Sang X, Diorio C, Lin S, Doillon C (2015) In vitro interactions between mammary fibroblasts (Hs 578Bst) and cancer epithelial cells (MCF-7) modulate aromatase, steroid sulfatase and 17β-hydroxysteroid dehydrogenases. Mol Cell Endocrinol 412:339–348. doi: 10.1016/j.mce.2015.05.032 CrossRefPubMedGoogle Scholar
- 11.McNamara K, Yoda T, Miki Y, Chanplakorn N, Wongwaisayawan S, Incharoen P, Kongdan Y, Wang L, Takagi K, Takagi M, Nakamura Y, Suzuki T, Nemoto N, Miyashita M, Tamaki K, Ishida T, Ohuchi N, Sasano H (2013) Androgenic pathway in triple negative invasive ductal tumors: Its correlation with tumor cell proliferation. Cancer Sci 104(5):639–646. doi: 10.1111/cas.12121 CrossRefPubMedGoogle Scholar
- 13.Miki Y, Suzuki T, Abe K, Suzuki S, Niikawa H, Iida S, Hata S, Akahira J, Mori K, Evans D, Kondo T, Yamada-Okabe H, Sasano H (2010) Intratumoral localization of aromatase and interaction between stromal and parenchymal cells in the non-small cell lung carcinoma microenvironment. Cancer Res 70(16):6659–6669. doi: 10.1158/0008-5472.CAN-09-4653 CrossRefPubMedGoogle Scholar
- 14.Lehmann B, Bauer J, Chen X, Sanders M, Chakravarthy A, Shyr Y, Pietenpol J (2011) Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. J Clin Invest 121(7):2750–2767. doi: 10.1172/JCI45014DS1 CrossRefPubMedPubMedCentralGoogle Scholar
- 15.Moore N, Buchanan G, Harris J, Selth L, Bianco-Miotto T, Hanson A, Birrell S, Butler L, Hickey T, Tilley W (2012) An androgen receptor mutation in the MDA-MB-453 cell line model of molecular apocrine breast cancer compromises receptor activity. Endocr-Related Cancer. 19(4):599–613. doi: 10.1530/ERC-12-0065 CrossRefGoogle Scholar
- 20.Cochrane D, Bernales S, Jacoben B, Cittelly D, Howe E, D’Amato N, Spoelstra N, Edgerton S, Jean A, Guerrero J, Gómez F, Medicherla S, Alfaro I, McCullagh E, Jedlicka P, Torkko K, Thor A, Elias A, Protter A, Richer J (2014) Role of the androgen receptor in breast cancer and preclinical analysis of enzalutamide. Breast Cancer Res 16(1):R7. doi: 10.1186/bcr3599 CrossRefPubMedPubMedCentralGoogle Scholar
- 27.Gritsko T, Williams A, Turkson J, Kaneko S, Bowman T, Huang M, Nam S, Eweis I, Diaz N, Sullivan D, Yoder S, Enkemann S, Eschrich S, Lee J, Beam C, Cheng J, Minton S, Muro-Cacho C, Jove R (2006) Persistent activation of Stat3 signaling induces survivin gene expression and confers resistance to apoptosis in human breast cancer cells. Clin Cancer Res 12(1):11–19. doi: 10.1158/1078-0432.CCR-04-1752 CrossRefPubMedGoogle Scholar
- 30.Bartholomeusz C, Gonzalez-angulo A, Liu P, Hayashi N, Lluch A, Ferrer-lozano J, Hortobagyi G (2012) High ERK protein expression levels correlate with shorter survival in triple-negative breast cancer patients. Oncologist 17(6):766–774. doi: 10.1634/theoncologist.2011-0377 CrossRefPubMedPubMedCentralGoogle Scholar
- 31.Eralp Y, Derin D, Ozluk Y, Yavuz E, Guney N, Saip P, Muslumanoglu M, Igci A, Kücücük S, Dincer M, Aydiner A, Topuz E (2008) MAPK overexpression is associated with anthracycline resistance and increased risk for recurrence in patients with triple-negative breast cancer. Ann Oncol 19(4):669–674. doi: 10.1093/annonc/mdm522 CrossRefPubMedGoogle Scholar
- 32.Kilvaer T, Khanehkenari M, Hellevik T, Al-Saad S, Paulsen E, Bremnes R, Busund L, Donnem T, Martinez I (2015) Cancer associated fibroblasts in stage I-IIIA NSCLC: prognostic impact and their correlations with tumor molecular markers. PLoS ONE 10(8):e0134965. doi: 10.1371/journal.pone.0134965 CrossRefPubMedPubMedCentralGoogle Scholar
- 34.McNamara K, Yoda T, Nurani A, Shibahara Y, Miki Y, Wang L, Nakamura Y, Suzuki K, Yang Y, Abe E, Hirakawa H, Suzuki T, Nemoto N, Miyashita M, Tamaki K, Ishida T, Brown K, Ohuchi N, Sasano H (2014) Androgenic pathways in the progression of triple-negative breast carcinoma: a comparison between aggressive and non-aggressive subtypes. Breast Cancer Res Tr 145(2):281–293. doi: 10.1007/s10549-014-2942-6 CrossRefGoogle Scholar
- 35.Takagi K, Miki Y, Nakagawa S, Hirakawa H, Onodera Y, Akahira J, Ishida T, Watanabe M, Kimizima I, Hayashi S, Sasano H, Suzuki T (2010) Increased intratumoral androgens in human breast carcinoma following aromatase inhibitor exemestane treatment. Endocr-Relat Cancer 17(2):415–430. doi: 10.1677/ERC-09-0257 CrossRefPubMedGoogle Scholar
- 36.McNamara K, Yoda T, Miki Y, Nakamura Y, Suzuki T, Nemoto N, Miyashita M, Nishimura R, Arima N, Tamaki K, Ishida T, Ohuchi N, Sasano H (2015) Androgen receptor and enzymes in lymph node metastasis and cancer reoccurrence in triple-negative breast cancer. Int J Biol Markers 30(2):e184–e189. doi: 10.5301/jbm.5000132 CrossRefPubMedGoogle Scholar
- 42.Gucalp A, Tolaney S, Isakoff S, Ingle J, Liu M, Carey L, Blackwell K, Rugo H, Nabell L, Forero A, Stearns V, Doane A, Danso M, Moynahan M, Momen L, Gonzalez J, Akhtar A, Giri D, Patil S, Feigin K, Hudis C, Traina T (2013) Phase II trial of bicalutamide in patients with androgen receptor-positive, estrogen receptor-negative metastatic breast cancer. Clin Cancer Res 19(19):5505–5512. doi: 10.1158/1078-0432.CCR-12-3327 CrossRefPubMedPubMedCentralGoogle Scholar
- 43.Barton V, D’Amato N, Gordon M, Lind H, Spoelstra N, Babbs B, Heinz R, Elias A, Jedlicka P, Jacobsen B, Richer J (2015) Multiple molecular subtypes of triple-negative breast cancer critically rely on androgen receptor and respond to enzalutamide In Vivo. Mol Cancer Ther 14(3):769–778. doi: 10.1158/1535-7163.MCT-14-0926 CrossRefPubMedPubMedCentralGoogle Scholar
- 45.Narayanan R, Ahn S, Cheney M, Yepuru M, Miller D, Steiner M, Dalton J (2014) Selective androgen receptor modulators (SARMs) negatively regulate triple-negative breast cancer growth and epithelial: mesenchymal stem cell signaling. PLoS ONE 9(7):e103202. doi: 10.1371/journal.pone.0103202 CrossRefPubMedPubMedCentralGoogle Scholar
- 49.Yoda T, McNamara K, Miki Y, Takagi M, Rai Y, Ohi Y, Sagara Y, Tamaki K, Hirakawa H, Ishida T, Suzuki T, Ohuchi N, Sasano H (2014) Intratumoral androgen metabolism and actions in invasive lobular carcinoma of the breast. Cancer Sci 105(11):1503–1509. doi: 10.1111/cas.12535 CrossRefPubMedPubMedCentralGoogle Scholar