Suberoylanilide hydroxamic acid upregulates reticulophagy receptor expression and promotes cell death in hepatocellular carcinoma cells

作     者：Jia-Yao Li Tian Tian Bing Han Ting Yang Yi-Xin Guo Jia-Yu Wu Yu-Si Chen Qin Yang Ru-Jia Xie 

作者机构：Guizhou Provincial Key Laboratory of Pathogenesis and Drug Research on Common Chronic DiseasesCollege of Basic Medical SciencesGuizhou Medical UniversityGuiyang 550025Guizhou ProvinceChina Department of PathophysiologyCollege of Basic Medical SciencesGuizhou Medical UniversityGuiyang 550025Guizhou ProvinceChina Department of Eugenic GeneticsGuiyang Maternal and Child Health Care HospitalGuiyang 550003Guizhou ProvinceChina 

出 版 物：《世界胃肠病学杂志:英文版》 (World Journal of Gastroenterology)

年 卷 期：2023年第29卷第34期

页      面：5038-5053页

核心收录：

学科分类：1006[医学-中西医结合] 1002[医学-临床医学] 10[医学] 100602[医学-中西医结合临床] 

基　　金：the National Natural Science Foundation of China,No.82260127 Guizhou Provincial Science and Technology Projects,No.Qiankehe Jichu-ZK365 and Qiankehe Jichu-ZK364 National Natural Science Foundation Cultivation Project of Guizhou Medical University,No.20NSP016 Guizhou Provincial Natural Science Foundation,No.4029 and4017 Science and Technology Foundation of Guizhou Provincial Health Commission,No.gzwjkj2019-1-102. 

主　　题：Suberoylanilide hydroxamic acid Histone H4 lysine-16 Reticulophagy Apoptosis Autophagic cell death Hepatocellular carcinoma 

摘      要：BACKGROUND Hepatocellular carcinoma(HCC)is a common clinical condition with a poor prognosis and few effective treatment options.Potent anticancer agents for treating HCC must be identified.Epigenetics plays an essential role in HCC tumorigenesis.Suberoylanilide hydroxamic acid(SAHA),the most common histone deacetylase inhibitor agent,triggers many forms of cell death in HCC.However,the underlying mechanism of action remains unclear.Family with sequence similarity 134 member B(FAM134B)-induced reticulophagy,a selective autophagic pathway,participates in the decision of cell fate and exhibits anticancer activity.This study focused on the relationship between FAM134B-induced reticulophagy and SAHA-mediated cell death.AIM To elucidate potential roles and underlying molecular mechanisms of reticulophagy in SAHA-induced HCC cell death.METHODS The viability,apoptosis,cell cycle,migration,and invasion of SAHA-treated Huh7 and MHCC97L cells were measured.Proteins related to the reticulophagy pathway,mitochondria-endoplasmic reticulum(ER)contact sites,intrinsic mitochondrial apoptosis,and histone acetylation were quantified using western blotting.ER and lysosome colocalization,and mitochondrial Ca^(2+)levels were characterized via confocal microscopy.The level of cell death was evaluated through Hoechst 33342 staining and propidium iodide colocalization.Chromatin immunoprecipitation was used to verify histone H4 lysine-16 acetylation in the FAM134B promoter region.RESULTS After SAHA treatment,the proliferation of Huh7 and MHCC97L cells was significantly inhibited,and the migration and invasion abilities were greatly blocked in vitro.This promoted apoptosis and caused G1 phase cells to increase in a concentration-dependent manner.Following treatment with SAHA,ER-phagy was activated,thereby triggering autophagy-mediated cell death of HCC cells in vitro.Western blotting and chromatin immunoprecipitation assays confirmed that SAHA regulated FAM134B expression by enhancing the histone H4 lysine-16 acetylation in the FAM134B promoter region.Further,SAHA disturbed the Ca^(2+)homeostasis and upregulated the level of autocrine motility factor receptor and proteins related to mitochondria-endoplasmic reticulum contact sites in HCC cells.Additionally,SAHA decreased the mitochondrial membrane potential levels,thereby accelerating the activation of the reticulophagy-mediated mitochondrial apoptosis pathway and promoting HCC cell death in vitro.CONCLUSION SAHA stimulates FAM134B-mediated ER-phagy to synergistically enhance the mitochondrial apoptotic pathway,thereby enhancing HCC cell death.