目的本研究聚焦于组蛋白乙酰化调控因子影响胰腺癌耐药中的机制。方法从癌症基因图谱(the cancer genome atlas,TCGA)数据库获取胰腺癌患者的RNA测序数据和临床病理信息,进行共识聚类分析和基因集富集分析。在胰腺癌细胞系PANC-1中敲减...目的本研究聚焦于组蛋白乙酰化调控因子影响胰腺癌耐药中的机制。方法从癌症基因图谱(the cancer genome atlas,TCGA)数据库获取胰腺癌患者的RNA测序数据和临床病理信息,进行共识聚类分析和基因集富集分析。在胰腺癌细胞系PANC-1中敲减和过表达组蛋白去乙酰化酶5(histone deacetylase 5,HDAC5)和特殊富含AT序列结合蛋白1(special AT rich sequence binding protein 1,SATB1),检测细胞活力和基因表达。构建裸鼠皮下移植瘤模型,观察HDAC5和SATB1表达水平对肿瘤生长和化疗敏感性的影响。结果基于组蛋白乙酰转移酶(histone acetyltransferase,HAT)和HDAC家族基因表达水平,将胰腺癌样本聚类为C1和C2两个亚型,C1亚型预后较差且化疗耐药相关通路显著富集。HDAC5在化疗耐药胰腺癌细胞中低表达,与患者预后差相关,且其表达水平与其启动子区甲基化水平呈负相关。SATB1在胰腺癌中的表达与化疗耐药性相关,主要富集于细胞周期、DNA复制和修复等通路。筛选出TRMU、THOP1、DLGAP1、LIPH、PRR15、KLK11和EHF等HDAC5/SATB1下游基因,其表达水平与胰腺癌患者预后和化疗敏感性密切相关。体内外实验证实,HDAC5低表达增强胰腺癌细胞对化疗药物的耐药性,而SATB1低表达则降低耐药性。结论HDAC5表达与胰腺癌患者预后呈正相关,其低表达与胰腺癌化疗耐药密切相关。HDAC5/SATB1轴在调控胰腺癌化疗耐药中起重要作用,为胰腺癌治疗提供潜在靶点。展开更多
目的:观察黄芪甲苷(AS-IV)通过调控蛋白激酶D1(PKD1)-组蛋白脱乙酰酶5(HDAC5)-血管内皮生长因子(VEGF)信号通路促心肌梗死大鼠血管新生的作用并分析其可能的作用机制。方法:采用经典的左冠状动脉前降支结扎术复制心肌梗死模型后,将大鼠...目的:观察黄芪甲苷(AS-IV)通过调控蛋白激酶D1(PKD1)-组蛋白脱乙酰酶5(HDAC5)-血管内皮生长因子(VEGF)信号通路促心肌梗死大鼠血管新生的作用并分析其可能的作用机制。方法:采用经典的左冠状动脉前降支结扎术复制心肌梗死模型后,将大鼠分成模型组、AS-IV治疗组和AS-IV+CID755673(PKD1阻断剂)组,另设假手术对照组和二甲基亚砜(DMSO)对照组,均采用尾静脉注射的给药方式。4周后处死大鼠,应用HE染色和Masson染色分析左心室心肌组织病理学变化;应用RT-PCR分析心肌组织中PKD1、HDAC5和VEGF m RNA的表达;免疫组化及Western blot法分析心肌组织中PKD1、VEGF及HDAC5蛋白的表达。结果:组织病理学分析表明,假手术组和DMSO组大鼠心肌组织形态正常,而模型组大鼠心肌组织形态紊乱,心肌细胞坏死和纤维化明显;AS-IV治疗后,心肌组织形态改善明显,且新生的血管数量明显增多;AS-IV+CID755673处理后大鼠心肌组织形态再次趋向紊乱,坏死细胞增多,部分血管闭合。模型组心肌组织中PKD1、HDAC5和VEGF的m RNA和蛋白表达明显低于假手术组和DMSO组(P<0.05),而AS-IV组显著高于模型组(P<0.01),AS-IV+CID755673组明显低于AS-IV组(P<0.05)。结论:AS-IV可部分通过调控PKD1-HDAC5-VEGF信号通路发挥促大鼠心肌梗死后心肌组织血管新生的作用。展开更多
The dysregulation of exosomal microRNAs(miRNAs)plays a crucial role in the development and progression of cancer.This study investigated the role of a newly identified serum exosomal miRNA miR-4256 in gastric cancer(G...The dysregulation of exosomal microRNAs(miRNAs)plays a crucial role in the development and progression of cancer.This study investigated the role of a newly identified serum exosomal miRNA miR-4256 in gastric cancer(GC)and the underlying mechanisms.The differentially expressed miRNAs were firstly identified in serum exosomes of GC patients and healthy individuals using next-generation sequencing and bioinformatics.Next,the expression of serum exosomal miR-4256 was analyzed in GC cells and GC tissues,and the role of miR-4256 in GC was investigated by in vitro and in vivo experiments.Then,the effect of miR-4256 on its downstream target genes HDAC5/p16^(INK4a) was studied in GC cells,and the underlying mechanisms were evaluated using dual luciferase reporter assay and Chromatin Immunoprecipitation(ChIP).Additionally,the role of the miR-4256/HDAC5/p16^(INK4a) axis in GC was studied using in vitro and in vivo experiments.Finally,the upstream regulators SMAD2/p300 that regulate miR-4256 expression and their role in GC were explored using in vitro experiments.miR-4256 was the most significantly upregulated miRNA and was overexpressed in GC cell lines and GC tissues;in vitro and in vivo results showed that miR-4256 promoted GC growth and progression.Mechanistically,miR-4256 enhanced HDAC5 expression by targeting the promoter of the HDAC5 gene in GC cells,and then restrained the expression of p16^(INK4a) through the epigenetic modulation of HDAC5 at the p16INK4a promoter.Furthermore,miR-4256 overexpression was positively regulated by the SMAD2/p300 complex in GC cells.Our data indicate that miR-4256 functions as an oncogene in GC via the SMAD2/miR-4256/HDAC5/p16^(INK4a) axis,which participates in GC progression and provides novel therapeutic and prognostic biomarkers for GC.展开更多
文摘目的本研究聚焦于组蛋白乙酰化调控因子影响胰腺癌耐药中的机制。方法从癌症基因图谱(the cancer genome atlas,TCGA)数据库获取胰腺癌患者的RNA测序数据和临床病理信息,进行共识聚类分析和基因集富集分析。在胰腺癌细胞系PANC-1中敲减和过表达组蛋白去乙酰化酶5(histone deacetylase 5,HDAC5)和特殊富含AT序列结合蛋白1(special AT rich sequence binding protein 1,SATB1),检测细胞活力和基因表达。构建裸鼠皮下移植瘤模型,观察HDAC5和SATB1表达水平对肿瘤生长和化疗敏感性的影响。结果基于组蛋白乙酰转移酶(histone acetyltransferase,HAT)和HDAC家族基因表达水平,将胰腺癌样本聚类为C1和C2两个亚型,C1亚型预后较差且化疗耐药相关通路显著富集。HDAC5在化疗耐药胰腺癌细胞中低表达,与患者预后差相关,且其表达水平与其启动子区甲基化水平呈负相关。SATB1在胰腺癌中的表达与化疗耐药性相关,主要富集于细胞周期、DNA复制和修复等通路。筛选出TRMU、THOP1、DLGAP1、LIPH、PRR15、KLK11和EHF等HDAC5/SATB1下游基因,其表达水平与胰腺癌患者预后和化疗敏感性密切相关。体内外实验证实,HDAC5低表达增强胰腺癌细胞对化疗药物的耐药性,而SATB1低表达则降低耐药性。结论HDAC5表达与胰腺癌患者预后呈正相关,其低表达与胰腺癌化疗耐药密切相关。HDAC5/SATB1轴在调控胰腺癌化疗耐药中起重要作用,为胰腺癌治疗提供潜在靶点。
文摘目的:观察黄芪甲苷(AS-IV)通过调控蛋白激酶D1(PKD1)-组蛋白脱乙酰酶5(HDAC5)-血管内皮生长因子(VEGF)信号通路促心肌梗死大鼠血管新生的作用并分析其可能的作用机制。方法:采用经典的左冠状动脉前降支结扎术复制心肌梗死模型后,将大鼠分成模型组、AS-IV治疗组和AS-IV+CID755673(PKD1阻断剂)组,另设假手术对照组和二甲基亚砜(DMSO)对照组,均采用尾静脉注射的给药方式。4周后处死大鼠,应用HE染色和Masson染色分析左心室心肌组织病理学变化;应用RT-PCR分析心肌组织中PKD1、HDAC5和VEGF m RNA的表达;免疫组化及Western blot法分析心肌组织中PKD1、VEGF及HDAC5蛋白的表达。结果:组织病理学分析表明,假手术组和DMSO组大鼠心肌组织形态正常,而模型组大鼠心肌组织形态紊乱,心肌细胞坏死和纤维化明显;AS-IV治疗后,心肌组织形态改善明显,且新生的血管数量明显增多;AS-IV+CID755673处理后大鼠心肌组织形态再次趋向紊乱,坏死细胞增多,部分血管闭合。模型组心肌组织中PKD1、HDAC5和VEGF的m RNA和蛋白表达明显低于假手术组和DMSO组(P<0.05),而AS-IV组显著高于模型组(P<0.01),AS-IV+CID755673组明显低于AS-IV组(P<0.05)。结论:AS-IV可部分通过调控PKD1-HDAC5-VEGF信号通路发挥促大鼠心肌梗死后心肌组织血管新生的作用。
基金The studies involving human participants were approved by The First Affiliated Hospital of Jinan University Ethics Committee(KY-2021-095)The participants provided their written informed consent to participate in this study+1 种基金Animalinvolved experimental protocols were compliance with guidelines and licensesapproved by the Laboratory Animal Center of Jinan University(20220225-65).
文摘The dysregulation of exosomal microRNAs(miRNAs)plays a crucial role in the development and progression of cancer.This study investigated the role of a newly identified serum exosomal miRNA miR-4256 in gastric cancer(GC)and the underlying mechanisms.The differentially expressed miRNAs were firstly identified in serum exosomes of GC patients and healthy individuals using next-generation sequencing and bioinformatics.Next,the expression of serum exosomal miR-4256 was analyzed in GC cells and GC tissues,and the role of miR-4256 in GC was investigated by in vitro and in vivo experiments.Then,the effect of miR-4256 on its downstream target genes HDAC5/p16^(INK4a) was studied in GC cells,and the underlying mechanisms were evaluated using dual luciferase reporter assay and Chromatin Immunoprecipitation(ChIP).Additionally,the role of the miR-4256/HDAC5/p16^(INK4a) axis in GC was studied using in vitro and in vivo experiments.Finally,the upstream regulators SMAD2/p300 that regulate miR-4256 expression and their role in GC were explored using in vitro experiments.miR-4256 was the most significantly upregulated miRNA and was overexpressed in GC cell lines and GC tissues;in vitro and in vivo results showed that miR-4256 promoted GC growth and progression.Mechanistically,miR-4256 enhanced HDAC5 expression by targeting the promoter of the HDAC5 gene in GC cells,and then restrained the expression of p16^(INK4a) through the epigenetic modulation of HDAC5 at the p16INK4a promoter.Furthermore,miR-4256 overexpression was positively regulated by the SMAD2/p300 complex in GC cells.Our data indicate that miR-4256 functions as an oncogene in GC via the SMAD2/miR-4256/HDAC5/p16^(INK4a) axis,which participates in GC progression and provides novel therapeutic and prognostic biomarkers for GC.
