Estrogen regulates Hippo signaling via GPER in breast cancer
Xin Zhou, Shuyang Wang, Zhen Wang, Xu Feng, Peng Liu, Xian-Bo Lv, Fulong Li, Fa-Xing Yu, Yiping Sun, Haixin Yuan, Hongguang Zhu, Yue Xiong, Qun-Ying Lei, Kun-Liang Guan
Xin Zhou, Shuyang Wang, Zhen Wang, Xu Feng, Peng Liu, Xian-Bo Lv, Fulong Li, Fa-Xing Yu, Yiping Sun, Haixin Yuan, Hongguang Zhu, Yue Xiong, Qun-Ying Lei, Kun-Liang Guan
View: Text | PDF
Research Article Oncology

Estrogen regulates Hippo signaling via GPER in breast cancer

Abstract

The G protein–coupled estrogen receptor (GPER) mediates both the genomic and nongenomic effects of estrogen and has been implicated in breast cancer development. Here, we compared GPER expression in cancerous tissue and adjacent normal tissue in patients with invasive ductal carcinoma (IDC) of the breast and determined that GPER is highly upregulated in cancerous cells. Additionally, our studies revealed that GPER stimulation activates yes-associated protein 1 (YAP) and transcriptional coactivator with a PDZ-binding domain (TAZ), 2 homologous transcription coactivators and key effectors of the Hippo tumor suppressor pathway, via the Gαq-11, PLCβ/PKC, and Rho/ROCK signaling pathways. TAZ was required for GPER-induced gene transcription, breast cancer cell proliferation and migration, and tumor growth. Moreover, TAZ expression positively correlated with GPER expression in human IDC specimens. Together, our results suggest that the Hippo/YAP/TAZ pathway is a key downstream signaling branch of GPER and plays a critical role in breast tumorigenesis.

Authors

Xin Zhou, Shuyang Wang, Zhen Wang, Xu Feng, Peng Liu, Xian-Bo Lv, Fulong Li, Fa-Xing Yu, Yiping Sun, Haixin Yuan, Hongguang Zhu, Yue Xiong, Qun-Ying Lei, Kun-Liang Guan

×

Figure 2

Stimulation of GPER activates YAP/TAZ.

Options: View larger image (or click on image) Download as PowerPoint
Stimulation of GPER activates YAP/TAZ. (A) Activation of GPER induced YA...
(A) Activation of GPER induced YAP dephosphorylation. Serum-starved SK-BR-3 cells were stimulated with 100 nM β-estradiol (E2), 200 nM OHT, or 100 nM G1. Immunoblotting was performed. (B) GPER activation led to TAZ dephosphorylation and accumulation. ZR-75-30 cells were serum starved and stimulated with 100 nM G1. TAZ phosphorylation was assessed by phos-tag gels. TAZ quantification is summarized in Supplemental Table 2. (C and D) GPER mediated TAZ activation. GPER was inhibited either by shRNAs (C) or 500 nM G15 (D), and ZR-75-30 cells were treated and immunoblotted as indicated. (E) G1 stimulated TAZ nuclear localization via GPER. Serum-starved ZR-75-30 cells were treated with G1 and/or G15, and immunofluorescence staining for TAZ was performed. Quantifications of TAZ subcellular localization from 100 randomly selected cells are shown. C, cytoplasmic; N, nuclear. (F) G1 enhanced TAZ interaction with TEAD1 but inhibited TAZ interaction with 14-3-30. Serum-starved ZR-75-30 cells were treated with G1 for 2 hours and then subjected to immunoprecipitation with TAZ antibody. The coimmunoprecipitated TEAD1 and 14-3-30 were detected. (G) Activation of GPER increased the expression of the YAP/TAZ target gene CTGF. Serum-starved SK-BR-3 or ZR-75-30 cells were treated with G1 or OHT as indicated. The quantification of CTGF expression is shown. (H) G1 stimulated TAZ and YAP nuclear localization in mammary glands. Mice were injected with G1 as described in the Supplemental Methods and stained for TAZ and YAP. Scale bars: 50 μm. Original magnification, ×400 (insets). The percentage of nuclear TAZ and YAP was analyzed by Student’s t test. Horizontal lines represent the median; the top and bottom of the boxes represent the 75th and 25th percentiles. Data are representative of at least 3 independent experiments. p-YAP, phosphorylated YAP.