BACKGROUND Endoplasmic reticulum(ER)stress-related hepatocyte apoptosis is responsible for multiple hepatic diseases.Previous studies have revealed that endoplasmic reticulophagy(ER-phagy)promotes the selective cleara...BACKGROUND Endoplasmic reticulum(ER)stress-related hepatocyte apoptosis is responsible for multiple hepatic diseases.Previous studies have revealed that endoplasmic reticulophagy(ER-phagy)promotes the selective clearance of damaged ER fragments during ER stress,playing a crucial role in maintaining ER homeostasis and inhibiting apoptosis.Family with sequence similarity 134 member B(FAM134B)is a receptor involved in ER-phagy that can form a complex with calnexin(CNX)and microtubule-associated protein 1 light chain 3(LC3).The complex can mediate the selective isolation of ER fragments to attenuate hepatocyte apoptosis.However,the precise regulatory mechanisms remain unclear.AIM To elucidate the effect of FAM134B-mediated ER-phagy on ER stress-induced apoptosis in buffalo rat liver 3A(BRL-3A)rat hepatocytes and the potential regulatory mechanisms.METHODS ER stress-related hepatocyte apoptosis was induced using dithiothreitol(DTT).Proteins related to ER stress and autophagy were measured with western blotting.Protein complex interactions with FAM134B were isolated by co-immunoprecipitation.ER-phagy was evaluated in immunofluorescence experiments.Cell cycle distribution and apoptosis were measured by flow cytometry.Mitochondrial Ca^(2+) levels were evaluated by the co-localization of intracellular Ca^(2+)-tracker and Mitotracker.The small interfering RNA against FAM134B was used to knockdown FAM134B in BRL-3A cells.RESULTS ER stress-related and autophagy-related proteins in BRL-3A cells were elevated by both short and long-term DTT treatment.Furthermore,co-immunoprecipitation confirmed an interaction between FAM134B,CNX,FAM134B,and LC3 in BRL-3A cells.Immunofluorescence assays revealed that autolysosomes significantly decreased following short-term DTT treatment,but increased after long-term treatment.Mitochondrial Ca2+levels and apoptotic rates were dramatically elevated,and more cells were arrested in the G1 stage after short-term DTT treatment;however,these decreased 48 h later.Moreover,FAM134B downregulation accelerated mitochondrial apoptotic pathway activation and aggravated hepatocyte apoptosis under ER stress.CONCLUSION FAM134B-mediated ER-phagy attenuates hepatocyte apoptosis by suppressing the mitochondrial apoptotic pathway.Our findings provide new evidence highlighting the importance of FAM134Bmediated ER-phagy in attenuating hepatocyte apoptosis.展开更多
目的采用多种生物信息学分析工具和免疫组织化学染色,明确序列相似性20家族成员A(Family with sequence similarity 20 member A,FAM20A)在甲状腺癌(Thyroid carcinoma,TC)中的表达及临床诊断意义。方法通过癌症基因组图谱(TCGA)数据库...目的采用多种生物信息学分析工具和免疫组织化学染色,明确序列相似性20家族成员A(Family with sequence similarity 20 member A,FAM20A)在甲状腺癌(Thyroid carcinoma,TC)中的表达及临床诊断意义。方法通过癌症基因组图谱(TCGA)数据库获取甲状腺癌相关数据,分析FAM20A在甲状腺癌组织和正常组织之间的表达差异,绘制ROC曲线计算AUC值,评估FAM20A对甲状腺癌的诊断效能。利用GEO数据库下载GSE205733甲状腺癌数据集,分析FAM20A基因在甲状腺癌组织中的表达水平,并进行基因富集分析(Gene set enrichment analysis,GSEA);利用在线数据库STRING分析FAM20A的蛋白相互作用网络。同时收集2022年3月—2023年2月期间于首都医科大学附属北京潞河医院甲状腺外科行甲状腺结节手术的29例患者病理组织样本,进行病理组织石蜡切片和免疫组织化学染色,验证比较甲状腺癌及良性甲状腺结节样本中FAM20A的表达水平。结果TCGA数据库结果表明,与正常甲状腺组织相比,FAM20A在甲状腺癌组织中的表达显著升高(P<0.001)。FAM20A表达水平与甲状腺癌临床TNM分期和病理分型相关(P<0.05)。FAM20A对于甲状腺癌的诊断效能显著(AUC=0.828)。GSE205733数据分析显示与正常组织相比,FAM20A在甲状腺癌组织中表达显著增加(P<0.001)。GSEA富集分析结果显示FAM20A主要与P53和白介素等信号传导通路相关。STRING数据库分析显示与FAM20A相互作用的蛋白主要有PEAK,FAM20C和ENAM等。免疫组织化学染色结果显示FAM20A在甲状腺癌组织中表达显著增加(P<0.05)。结论FAM20A基因在甲状腺癌组织中高表达,与患者临床TNM分期和病理分型相关,可作为诊断甲状腺癌的潜在分子标记物,且与P53和白介素等信号传导通路相关,为研究甲状腺癌进展分子机制及靶向药物的开发提供理论依据。展开更多
目的研究FAM134B在肝癌组织中的表达情况,探讨其表达差异与临床病理特征以及预后生存的关系。方法挖掘Oncomine、Human Protein Atlas、TCGA和Kaplan Meier-Plotter数据库,分析FAM134B在肝癌组织中的表达,并对表达量与临床病理特征以及...目的研究FAM134B在肝癌组织中的表达情况,探讨其表达差异与临床病理特征以及预后生存的关系。方法挖掘Oncomine、Human Protein Atlas、TCGA和Kaplan Meier-Plotter数据库,分析FAM134B在肝癌组织中的表达,并对表达量与临床病理特征以及预后生存的关系进行探讨。计数资料组间比较采用χ^2检验。FAM134B表达与肝癌预后的关系采用Kaplan-Meier模型分析。结果正常组织中,FAM134B在肾脏中表达量最高,肝脏居第9位,肿瘤组织中,其在肾癌组织表达最高,肝癌第11位。共147项有关FAM134B表达差异的研究结果差异具有统计学意义,各种癌症中其表达差异不一致。其在肝癌组织中表达量较正常肝组织中低。FAM134B蛋白定位于内质网,其表达量与肝癌患者发病年龄、肝硬化、AFP、人种、分化程度有相关性(P值均<0.05)。FAM134B表达量与肝癌患者总生存率有关[风险比(HR)=0.67,95%可信区间(95%CI):0.47~0.95,P=0.026],且对亚洲人种预后有显著影响(HR=0.40,95%CI:0.22~0.74,P=0.003),但对白色人种影响不显著(HR=0.65,95%CI:0.40~1.05,P=0.076)。结论FAM134B在不同癌症中表达不一致,在肝癌中低表达,蛋白定位于内质网,其低表达与肝癌部分恶性表型相关,且与患者总体生存率低有关。展开更多
序列相似家族172成员A(family with sequence similarity 172 member A,FAM172A)已被证实与多种肿瘤的发生和恶性转化有关,但是FAM172A在肝细胞癌(hepatocellular carcinoma,HCC)中的作用尚不完全清楚。本研究分析了肿瘤与癌症基因组图...序列相似家族172成员A(family with sequence similarity 172 member A,FAM172A)已被证实与多种肿瘤的发生和恶性转化有关,但是FAM172A在肝细胞癌(hepatocellular carcinoma,HCC)中的作用尚不完全清楚。本研究分析了肿瘤与癌症基因组图谱数据库中50例正常肝组织和50例肝癌组织信息,结合实时荧光定量PCR和蛋白质免疫印迹结果,发现肝癌组织中FAM172A的表达高于正常肝组织。利用短发夹RNA(short hairpin RNA,shRNA)将肝癌细胞系中FAM172A进行敲低,细胞活力检测和细胞克隆形成实验结果表明,FAM172A的敲低显著抑制了细胞的增殖(P<0.01)。此外糖酵解压力测试和Western印迹结果显示,敲低FAM172A可以有效抑制肝癌细胞的糖酵解能力,糖酵解酶己糖激酶2(hexokinase 2,HK2)、肝内磷酸果糖激酶(phosphofructokinaseliver,PFKL)、乳酸脱氢酶A(lactate dehydrogenase A,LDHA)及其转录因子c-Myc的表达也明显下调(P<0.001)。过表达c-Myc能够解除敲低FAM172A导致的细胞增殖抑制。综上所述,本文揭示了敲低FAM172A抑制肝癌细胞增殖以及发现FAM172A在肝癌中调控有氧糖酵解的新功能。展开更多
目的探究序列相似家族111成员A(family with sequence similarity 111 member A,FAM111A)、FAM111B在泛癌中的肿瘤预后、肿瘤免疫及抗癌药物敏感性。方法在癌症基因体图谱(the cancer genome atlas,TCGA)数据库下载和整理FAM111A和FAM1...目的探究序列相似家族111成员A(family with sequence similarity 111 member A,FAM111A)、FAM111B在泛癌中的肿瘤预后、肿瘤免疫及抗癌药物敏感性。方法在癌症基因体图谱(the cancer genome atlas,TCGA)数据库下载和整理FAM111A和FAM111B在33种肿瘤及11057例样本的mRNA表达水平及临床生存相关数据,下载UCSC Xena数据库中关于33种肿瘤干细胞评分相关数据,下载CellMiner数据库样本的基因表达与药敏结果的数据。对FAM111A与FAM111B在肿瘤中作用进行多方面研究。结果FAM111A和FAM111B的相关性较强(r=0.42,P<0.05),FAM111A和FAM111B在多种肿瘤中普遍高表达(P<0.05),且FAM111A和FAM111B可以预测多种肿瘤患者的生存率(P<0.05)。泛癌免疫亚型分析显示,FAM111A和FAM111B在6种肿瘤免疫亚型显著表达(P<0.001)。FAM111A和FAM111B的表达与免疫评分、间质评分及总评分呈负相关(P<0.05),FAM111A和FAM111B的表达与肿瘤干细胞分化程度呈正相关(P<0.05)。抗癌药物敏感性的分析显示,FAM111A与奈拉滨(Nelarabine)等药物敏感性呈正相关(P<0.05),FAM111基因与卡博替尼(Cabozantinib)等药物敏感性呈负相关(P<0.05)。结论FAM111A和FAM111B在多种肿瘤中有表达差异,并且对生存预后有预测价值,它们在肿瘤免疫微环境、干细胞评分和抗癌药物敏感性方面的研究结果为肿瘤治疗及诊断提供了方向。展开更多
基金Supported by National Natural Science Foundation of China,No.81560105Science and Technology Foundation of Guizhou Province,No.Qiankehe Jichu-ZK[2021]365,and No.Qiankehe Pingtai Rencai[2019]5801National Natural Science Foundation Cultivation Project of Guizhou Medical University,No.20NSP016。
文摘BACKGROUND Endoplasmic reticulum(ER)stress-related hepatocyte apoptosis is responsible for multiple hepatic diseases.Previous studies have revealed that endoplasmic reticulophagy(ER-phagy)promotes the selective clearance of damaged ER fragments during ER stress,playing a crucial role in maintaining ER homeostasis and inhibiting apoptosis.Family with sequence similarity 134 member B(FAM134B)is a receptor involved in ER-phagy that can form a complex with calnexin(CNX)and microtubule-associated protein 1 light chain 3(LC3).The complex can mediate the selective isolation of ER fragments to attenuate hepatocyte apoptosis.However,the precise regulatory mechanisms remain unclear.AIM To elucidate the effect of FAM134B-mediated ER-phagy on ER stress-induced apoptosis in buffalo rat liver 3A(BRL-3A)rat hepatocytes and the potential regulatory mechanisms.METHODS ER stress-related hepatocyte apoptosis was induced using dithiothreitol(DTT).Proteins related to ER stress and autophagy were measured with western blotting.Protein complex interactions with FAM134B were isolated by co-immunoprecipitation.ER-phagy was evaluated in immunofluorescence experiments.Cell cycle distribution and apoptosis were measured by flow cytometry.Mitochondrial Ca^(2+) levels were evaluated by the co-localization of intracellular Ca^(2+)-tracker and Mitotracker.The small interfering RNA against FAM134B was used to knockdown FAM134B in BRL-3A cells.RESULTS ER stress-related and autophagy-related proteins in BRL-3A cells were elevated by both short and long-term DTT treatment.Furthermore,co-immunoprecipitation confirmed an interaction between FAM134B,CNX,FAM134B,and LC3 in BRL-3A cells.Immunofluorescence assays revealed that autolysosomes significantly decreased following short-term DTT treatment,but increased after long-term treatment.Mitochondrial Ca2+levels and apoptotic rates were dramatically elevated,and more cells were arrested in the G1 stage after short-term DTT treatment;however,these decreased 48 h later.Moreover,FAM134B downregulation accelerated mitochondrial apoptotic pathway activation and aggravated hepatocyte apoptosis under ER stress.CONCLUSION FAM134B-mediated ER-phagy attenuates hepatocyte apoptosis by suppressing the mitochondrial apoptotic pathway.Our findings provide new evidence highlighting the importance of FAM134Bmediated ER-phagy in attenuating hepatocyte apoptosis.
文摘目的采用多种生物信息学分析工具和免疫组织化学染色,明确序列相似性20家族成员A(Family with sequence similarity 20 member A,FAM20A)在甲状腺癌(Thyroid carcinoma,TC)中的表达及临床诊断意义。方法通过癌症基因组图谱(TCGA)数据库获取甲状腺癌相关数据,分析FAM20A在甲状腺癌组织和正常组织之间的表达差异,绘制ROC曲线计算AUC值,评估FAM20A对甲状腺癌的诊断效能。利用GEO数据库下载GSE205733甲状腺癌数据集,分析FAM20A基因在甲状腺癌组织中的表达水平,并进行基因富集分析(Gene set enrichment analysis,GSEA);利用在线数据库STRING分析FAM20A的蛋白相互作用网络。同时收集2022年3月—2023年2月期间于首都医科大学附属北京潞河医院甲状腺外科行甲状腺结节手术的29例患者病理组织样本,进行病理组织石蜡切片和免疫组织化学染色,验证比较甲状腺癌及良性甲状腺结节样本中FAM20A的表达水平。结果TCGA数据库结果表明,与正常甲状腺组织相比,FAM20A在甲状腺癌组织中的表达显著升高(P<0.001)。FAM20A表达水平与甲状腺癌临床TNM分期和病理分型相关(P<0.05)。FAM20A对于甲状腺癌的诊断效能显著(AUC=0.828)。GSE205733数据分析显示与正常组织相比,FAM20A在甲状腺癌组织中表达显著增加(P<0.001)。GSEA富集分析结果显示FAM20A主要与P53和白介素等信号传导通路相关。STRING数据库分析显示与FAM20A相互作用的蛋白主要有PEAK,FAM20C和ENAM等。免疫组织化学染色结果显示FAM20A在甲状腺癌组织中表达显著增加(P<0.05)。结论FAM20A基因在甲状腺癌组织中高表达,与患者临床TNM分期和病理分型相关,可作为诊断甲状腺癌的潜在分子标记物,且与P53和白介素等信号传导通路相关,为研究甲状腺癌进展分子机制及靶向药物的开发提供理论依据。
文摘目的研究FAM134B在肝癌组织中的表达情况,探讨其表达差异与临床病理特征以及预后生存的关系。方法挖掘Oncomine、Human Protein Atlas、TCGA和Kaplan Meier-Plotter数据库,分析FAM134B在肝癌组织中的表达,并对表达量与临床病理特征以及预后生存的关系进行探讨。计数资料组间比较采用χ^2检验。FAM134B表达与肝癌预后的关系采用Kaplan-Meier模型分析。结果正常组织中,FAM134B在肾脏中表达量最高,肝脏居第9位,肿瘤组织中,其在肾癌组织表达最高,肝癌第11位。共147项有关FAM134B表达差异的研究结果差异具有统计学意义,各种癌症中其表达差异不一致。其在肝癌组织中表达量较正常肝组织中低。FAM134B蛋白定位于内质网,其表达量与肝癌患者发病年龄、肝硬化、AFP、人种、分化程度有相关性(P值均<0.05)。FAM134B表达量与肝癌患者总生存率有关[风险比(HR)=0.67,95%可信区间(95%CI):0.47~0.95,P=0.026],且对亚洲人种预后有显著影响(HR=0.40,95%CI:0.22~0.74,P=0.003),但对白色人种影响不显著(HR=0.65,95%CI:0.40~1.05,P=0.076)。结论FAM134B在不同癌症中表达不一致,在肝癌中低表达,蛋白定位于内质网,其低表达与肝癌部分恶性表型相关,且与患者总体生存率低有关。
文摘目的探究序列相似家族111成员A(family with sequence similarity 111 member A,FAM111A)、FAM111B在泛癌中的肿瘤预后、肿瘤免疫及抗癌药物敏感性。方法在癌症基因体图谱(the cancer genome atlas,TCGA)数据库下载和整理FAM111A和FAM111B在33种肿瘤及11057例样本的mRNA表达水平及临床生存相关数据,下载UCSC Xena数据库中关于33种肿瘤干细胞评分相关数据,下载CellMiner数据库样本的基因表达与药敏结果的数据。对FAM111A与FAM111B在肿瘤中作用进行多方面研究。结果FAM111A和FAM111B的相关性较强(r=0.42,P<0.05),FAM111A和FAM111B在多种肿瘤中普遍高表达(P<0.05),且FAM111A和FAM111B可以预测多种肿瘤患者的生存率(P<0.05)。泛癌免疫亚型分析显示,FAM111A和FAM111B在6种肿瘤免疫亚型显著表达(P<0.001)。FAM111A和FAM111B的表达与免疫评分、间质评分及总评分呈负相关(P<0.05),FAM111A和FAM111B的表达与肿瘤干细胞分化程度呈正相关(P<0.05)。抗癌药物敏感性的分析显示,FAM111A与奈拉滨(Nelarabine)等药物敏感性呈正相关(P<0.05),FAM111基因与卡博替尼(Cabozantinib)等药物敏感性呈负相关(P<0.05)。结论FAM111A和FAM111B在多种肿瘤中有表达差异,并且对生存预后有预测价值,它们在肿瘤免疫微环境、干细胞评分和抗癌药物敏感性方面的研究结果为肿瘤治疗及诊断提供了方向。
