P300/CBP-associated factor (PCAF)-mediated acetylation of Fascin at lysine 471 inhibits its actin-bundling activity and tumor metastasis in esophageal cancer

Cancer Communications

article

Yin-Wei Cheng¹  Fa-Min Zeng¹  Da-Jia Li¹  Shao-Hong Wang⁴  Jian-Zhong He¹  Zhen-Chang Guo¹  Ping-Juan Nie¹  Zhi-Yong Wu⁴  Wen-Qi Shi¹  Bing Wen¹  Xiu-E Xu¹  Lian-Di Liao¹  Zhi-Mao Li¹  Jian-Yi Wu¹  Jun Zhan⁷  Hong-Quan Zhang⁷  Zhi-Jie Chang⁸  Kai Zhang⁶  Li-Yan Xu¹  En-Min Li¹ 

[1] The Key Laboratory of Molecular Biology for High Cancer Incidence Coastal Chaoshan Area, Department of Biochemistry and Molecular Biology, Shantou University Medical College;Institute of Basic Medical Science, Cancer Research Center, Shantou University Medical College;These authors contributed equally to this work.;Shantou Central Hospital, Affiliated Shantou Hospital of Sun Yat-sen University;Guangdong Provincial Key Laboratory of Infectious Diseases and Molecular Immunopathology, Institute of Oncologic Pathology, Shantou University Medical College;Tianjin Key Laboratory of Medical Epigenetics, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Tianjin Medical University;State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center;School of Medicine, Tsinghua University

关键词: acetylation;    actin-bundling;    esophageal cancer;    fascin;    filopodium formation;    P300/CBP-associated factor (PCAF);    tumor metastasis;   

DOI  :  10.1002/cac2.12221

学科分类：社会科学、人文和艺术（综合）

来源: Springer

【 摘 要 】

Background Fascin is crucial for cancer cell filopodium formation and tumor metastasis, and is functionally regulated by post-translational modifications. However, whether and how Fascin is regulated by acetylation remains unclear. This study explored the regulation of Fascin acetylation and its corresponding roles in filopodium formation and tumor metastasis. Methods Immunoprecipitation and glutathione-S-transferase pull-down assays were performed to examine the interaction between Fascin and acetyltransferase P300/CBP-associated factor (PCAF), and immunofluorescence was used to investigate their colocalization. An in vitro acetylation assay was performed to identify Fascin acetylation sites by using mass spectrometry. A specific antibody against acetylated Fascin was generated and used to detect the PCAF-mediated Fascin acetylation in esophageal squamous cell carcinoma (ESCC) cells using Western blotting by overexpressing and knocking down PCAF expression. An in vitro cell migration assay was performed, and a xenograft model was established to study in vivo tumor metastasis. Live-cell imaging and fluorescence recovery after photobleaching were used to evaluate the function and dynamics of acetylated Fascin in filopodium formation. The clinical significance of acetylated Fascin and PCAF in ESCC was evaluated using immunohistochemistry. Results Fascin directly interacted and colocalized with PCAF in the cytoplasm and was acetylated at lysine 471 (K471) by PCAF. Using the specific anti-AcK471-Fascin antibody, Fascin was found to be acetylated in ESCC cells, and the acetylation level was consequently increased after PCAF overexpression and decreased after PCAF knockdown. Functionally, Fascin-K471 acetylation markedly suppressed in vitro ESCC cell migration and in vivo tumor metastasis, whereas Fascin-K471 deacetylation exhibited a potent oncogenic function. Moreover, Fascin-K471 acetylation reduced filopodial length and density, and lifespan of ESCC cells, while its deacetylation produced the opposite effect. In the filipodium shaft, K471-acetylated Fascin displayed rapid dynamic exchange, suggesting that it remained in its monomeric form owing to its weakened actin-bundling activity. Clinically, high levels of AcK471-Fascin in ESCC tissues were strongly associated with prolonged overall survival and disease-free survival of ESCC patients. Conclusions Fascin interacts directly with PCAF and is acetylated at lysine 471 in ESCC cells. Fascin-K471 acetylation suppressed ESCC cell migration and tumor metastasis by reducing filopodium formation through the impairment of its actin-bundling activity.

【 授权许可】

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