Inhibition of histone deacetylase 6 destabilizes ERK phosphorylation and suppresses cancer proliferation via modulation of the tubulin acetylation-GRP78 interaction

Journal of Biomedical Science

Research

Piriya Wongkongkathep¹  Naphat Chantaravisoot²  Pithi Chanvorachote³  Onsurang Wattanathamsan³  Varisa Pongrakhananon⁴  Sakkarin Kungsukool⁵  Chanida Vinayanuwattikun⁶  Paninee Chetprayoon⁷ 

[1] Center of Excellence in Systems Biology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;Center of Excellence in Systems Biology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences,, Chulalongkorn University, Bangkok, Thailand;Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences,, Chulalongkorn University, Bangkok, Thailand;Preclinical Toxicity and Efficacy Assessment of Medicines and Chemicals Research Cluster, Chulalongkorn University, Bangkok, Thailand;Department of Respiratory Medicine, Central Chest Institute of Thailand, Muang District, Nonthaburi, Thailand;Division of Medical Oncology, Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;Toxicology and Bio Evaluation Service Center, National Science and Technology Development Agency, Pathum Thani, Thailand;

关键词: Extracellular signal-regulated kinase (ERK);    Glucose-regulated protein 78 (GRP78);    Histone deacetylase 6 (HDAC6);    Lung cancer;    Tubulin acetylation;   

DOI  :  10.1186/s12929-023-00898-3

 received in 2022-05-18, accepted in 2023-01-06,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundThe leading cause of cancer-related mortality worldwide is lung cancer, and its clinical outcome and prognosis are still unsatisfactory. The understanding of potential molecular targets is necessary for clinical implications in precision diagnostic and/or therapeutic purposes. Histone deacetylase 6 (HDAC6), a major deacetylase enzyme, is a promising target for cancer therapy; however, the molecular mechanism regulating cancer pathogenesis is largely unknown.MethodsThe clinical relevance of HDAC6 expression levels and their correlation with the overall survival rate were analyzed based on the TCGA and GEO databases. HDAC6 expression in clinical samples obtained from lung cancer tissues and patient-derived primary lung cancer cells was evaluated using qRT–PCR and Western blot analysis. The potential regulatory mechanism of HDAC6 was identified by proteomic analysis and validated by immunoblotting, immunofluorescence, microtubule sedimentation, and immunoprecipitation-mass spectrometry (IP-MS) assays using a specific inhibitor of HDAC6, trichostatin A (TSA) and RNA interference to HDAC6 (siHDAC6). Lung cancer cell growth was assessed by an in vitro 2-dimensional (2D) cell proliferation assay and 3D tumor spheroid formation using patient-derived lung cancer cells.ResultsHDAC6 was upregulated in lung cancer specimens and significantly correlated with poor prognosis. Inhibition of HDAC6 by TSA and siHDAC6 caused downregulation of phosphorylated extracellular signal-regulated kinase (p-ERK), which was dependent on the tubulin acetylation status. Tubulin acetylation induced by TSA and siHDAC6 mediated the dissociation of p-ERK on microtubules, causing p-ERK destabilization. The proteomic analysis demonstrated that the molecular chaperone glucose-regulated protein 78 (GRP78) was an important scaffolder required for p-ERK localization on microtubules, and this phenomenon was significantly inhibited by either TSA, siHDAC6, or siGRP78. In addition, suppression of HDAC6 strongly attenuated an in vitro 2D lung cancer cell growth and an in vitro 3D patient derived-lung cancer spheroid growth.ConclusionsHDAC6 inhibition led to upregulate tubulin acetylation, causing GRP78-p-ERK dissociation from microtubules. As a result, p-ERK levels were decreased, and lung cancer cell growth was subsequently suppressed. This study reveals the intriguing role and molecular mechanism of HDAC6 as a tumor promoter, and its inhibition represents a promising approach for anticancer therapy.

【 授权许可】

CC BY   
© The Author(s) 2023

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