HIC1 deletion promotes breast cancer progression by activating tumor cell/fibroblast crosstalk
Yingying Wang, … , Jinsong Lu, Jianhua Wang
Yingying Wang, … , Jinsong Lu, Jianhua Wang
Published September 11, 2018
Citation Information: J Clin Invest. 2018;128(12):5235-5250. https://doi.org/10.1172/JCI99974.
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Research Article Cell biology Oncology Article has an altmetric score of 4

HIC1 deletion promotes breast cancer progression by activating tumor cell/fibroblast crosstalk

Abstract

Breast cancer (BrCa) is the malignant tumor that most seriously threatens female health; however, the molecular mechanism underlying its progression remains unclear. Here, we found that conditional deletion of hypermethylated in cancer 1 (HIC1) in the mouse mammary gland might contribute to premalignant transformation in the early stage of tumor formation. Moreover, the chemokine (C-X-C motif) ligand 14 (CXCL14) secreted by HIC1-deleted BrCa cells bound to its cognate receptor GPR85 on mammary fibroblasts in the microenvironment and was responsible for activating these fibroblasts via the ERK1/2, Akt, and neddylation pathways, whereas the activated fibroblasts promoted BrCa progression via the induction of epithelial-mesenchymal transition (EMT) by the C-C chemokine ligand 17 (CCL17)/CC chemokine receptor 4 (CCR4) axis. Finally, we confirmed that the HIC1-CXCL14-CCL17 loop was associated with the malignant progression of BrCa. Therefore, the crosstalk between HIC1-deleted BrCa cells and mammary fibroblasts might play a critical role in BrCa development. Exploring the progression of BrCa from the perspective of microenvironment will be beneficial for identifying the potential prognostic markers of breast tumor and providing more effective treatment strategies.

Authors

Yingying Wang, Xiaoling Weng, Luoyang Wang, Mingang Hao, Yue Li, Lidan Hou, Yu Liang, Tianqi Wu, Mengfei Yao, Guowen Lin, Yiwei Jiang, Guohui Fu, Zhaoyuan Hou, Xiangjun Meng, Jinsong Lu, Jianhua Wang

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Figure 5

GPR85 is a functional receptor for CXCL14 activity.

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GPR85 is a functional receptor for CXCL14 activity. (A) Confocal microsc...
(A) Confocal microscopy of NAF6 cells treated with 100 ng/ml biotin or biotin-CXCL14 at 4°C and stained with an antibody against Cy3-streptavidin. An isotype-matched IgG was used as a control. Cell nuclei were counterstained with DAPI. (B) Schematic of the procedure used to detect biotin-CXCL14–binding proteins using HuProt human proteome microarrays containing 18,583 affinity-purified N-terminally GST-tagged proteins. (C) Representative CXCL14-binding membrane proteins in the proteome microarrays. (D) Western blot validation using a streptavidin-agarose pull-down assay of proteome microarray determination that CXCL14 binds directly to GPR85. (E) Mobilization of [Ca2+]i in NAF6 cells that were transfected with control siRNA (NC) or GPR85-3 siRNA and then treated with 100 ng/ml HBSS, rhCXCL14, or rhCXCL12. The black arrows denote the times at which stimulation was initiated. (F) 125I-CXCL14 binding properties between 293T-NC and 293T-GPR85 cells. Data are shown as mean ± SD. n = 4 repetitions. **P < 0.01; ***P < 0.001, 2-tailed Student’s t test. (G) Binding assay with 10 nM 125I-CXCL14 in the presence or absence of increasing concentrations of unlabeled rhCXCL14, rhCXCL12, and rhCXCL3 for 293T cells that were transfected with GPR85. B, specific binding; T, total binding. (H) Knockdown of GPR85 expression by GPR85-3 siRNA in NAF6 cells in the presence or absence of 100 ng/ml rhCXCL14 for the indicated times (0, 30, and 60 minutes). Cell lysates were analyzed by Western blot with antibodies against p-Akt (Ser 473), Akt, p-ERK1/2, ERK1/2, and GAPDH. (I) Knockdown of GPR85 expression by GPR85-3 siRNA in NAF6 cells treated with rhCXCL14 at various concentrations (0–100 ng/ml) for 4 days. Cell lysates were analyzed by Western blot with antibodies against α-SMA, FAP, PDGFRα, and GAPDH. (J) NAF6 cells were transfected with control siRNA (NC) or GPR85-3 siRNA and then cocultured with MCF7Ctrl or MCF7sgHIC1 cells, respectively, for 4 days. NAF6 cell lysates were analyzed by Western blot with antibodies against α-SMA, FAP, PDGFRα, p-Akt (Ser 473), Akt, p-ERK1/2, ERK1/2, and GAPDH.