ZMYND8 acetylation mediates HIF-dependent breast cancer progression and metastasis
Yan Chen, … , Yingfei Wang, Weibo Luo
Yan Chen, … , Yingfei Wang, Weibo Luo
Published April 9, 2018
Citation Information: J Clin Invest. 2018;128(5):1937-1955. https://doi.org/10.1172/JCI95089.
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Research Article Oncology

ZMYND8 acetylation mediates HIF-dependent breast cancer progression and metastasis

Abstract

Altered epigenetic reprogramming contributes to breast cancer progression and metastasis. How the epigenetic reader mediates breast cancer progression remains poorly understood. Here, we showed that the epigenetic reader zinc finger MYND-type containing 8 (ZMYND8) is induced by HIF-1 and HIF-2 in breast cancer cells and also upregulated in human breast tumors, and is correlated with poor survival of patients with breast cancer. Genetic deletion of ZMYND8 decreases breast cancer cell colony formation, migration, and invasion in vitro, and inhibits breast tumor growth and metastasis to the lungs in mice. The ZMYND8’s oncogenic effect in breast cancer requires HIF-1 and HIF-2. We further showed that ZMYND8 interacts with HIF-1α and HIF-2α and enhances elongation of the global HIF-induced oncogenic genes by increasing recruitment of BRD4 and subsequent release of paused RNA polymerase II in breast cancer cells. ZMYND8 acetylation at lysines 1007 and 1034 by p300 is required for HIF activation and breast cancer progression and metastasis. These findings uncover a primary epigenetic mechanism of HIF activation and HIF-mediated breast cancer progression, and discover a possible molecular target for the diagnosis and treatment of breast cancer.

Authors

Yan Chen, Bo Zhang, Lei Bao, Lai Jin, Mingming Yang, Yan Peng, Ashwani Kumar, Jennifer E. Wang, Chenliang Wang, Xuan Zou, Chao Xing, Yingfei Wang, Weibo Luo

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Figure 12

Acetylation of ZMYND8 by p300 is necessary for HIF activation and breast tumor progression.

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Acetylation of ZMYND8 by p300 is necessary for HIF activation and breast...
(A and B) Acetylation of ZMYND8-V5 (A) and WT or mutant FLAG-ZMYND8 (B) in HEK293T cells treated with TSA or DMSO (–) for 6 hours (n = 3). (C) Co-IP assays of BRD4 and WT or mutant FLAG-ZMYND8 in transfected HEK293T cells (n = 3). (D and E) In vitro acetylation assays of WT or K1007/1034R FLAG-ZMYND8 by purified FLAG-p300, FLAG-PCAF, or FLAG-GCN5 (n = 3). (F) Co-IP assays of endogenous ZMYND8 and p300 in MCF-7 cells (n = 3). (G) Acetylation of endogenous ZMYND8 in SC and p300-KD MCF-7 cells (n = 3). (H) Co-IP assays of endogenous ZMYND8 and BRD4 in SC and p300-KD MCF-7 cells (n = 3). (I) RT-qPCR analysis of indicated mRNAs in ZMYND8-rescued MDA-MB-231 cells exposed to 20% or 1% O2 for 24 hours (mean ± SEM, n = 3). *P < 0.05, **P < 0.01, by 2-way ANOVA with Tukey’s t test. (J–L) Clonogenic assays (J), migration assays (K), and invasion assays (L) in ZMYND8-rescued MDA-MB-231 cells exposed to 20% or 1% O2 for 12 days (J), 16 hours (K), or 24 hours (L) (mean ± SEM, n = 3). *P < 0.05, ***P < 0.001, ****P < 0.0001, by 2-way ANOVA with Tukey’s t test. (M–P) Growth of ZMYND8-rescued MDA-MB-231 tumors in mice (M, mean ± SEM, n = 5). Endomucin-positive areas (N) and CC3-positive cell numbers (O) in tumors and lung metastasis (P) were quantified (mean ± SEM, n = 5). **P < 0.01, ***P < 0.001, ****P < 0.0001, by 2-way ANOVA with Tukey’s t test.