Background:Liver cancer is a malignancy with high morbidity and mortality rates.Serpin family E member 2(SERPINE2)has been reported to play a key role in the metastasis of many tumors.In this study,we aimed to investi...Background:Liver cancer is a malignancy with high morbidity and mortality rates.Serpin family E member 2(SERPINE2)has been reported to play a key role in the metastasis of many tumors.In this study,we aimed to investigate the potential mechanism of SERPINE2 in liver cancer metastasis.Methods:The Cancer Genome Atlas database(TCGA),including DNA methy-lation and transcriptome sequencing data,was utilized to identify the crucial oncogene associated with DNA methylation and cancer progression in liver can-cer.Data from the TCGA and RNA sequencing for 94 pairs of liver cancer tissues were used to explore the correlation between SERPINE2 expression and clin-ical parameters of patients.DNA methylation sequencing was used to detect the DNA methylation levels in liver cancer tissues and cells.RNA sequencing,cytokine assays,immunoprecipitation(IP)and mass spectrometry(MS)assays,protein stability assays,and ubiquitination assays were performed to explore the regulatory mechanism of SERPINE2 in liver cancer metastasis.Patient-derived xenografts and tumor organoid models were established to determine the role of SERPINE2 in the treatment of liver cancer using sorafenib.Results:Based on the public database screening,SERPINE2 was identified as a tumor promoter regulated by DNA methylation.SERPINE2 expression was significantly higher in liver cancer tissues and was associated with the dismal prognosis in patients with liver cancer.SERPINE2 promoted liver cancer metas-tasis by enhancing cell pseudopodia formation,cell adhesion,cancer-associated fibroblast activation,extracellular matrix remodeling,and angiogenesis.IP/MS assays confirmed that SERPINE2 activated epidermal growth factor receptor(EGFR)and its downstream signaling pathways by interacting with EGFR.Mechanistically,SERPINE2 inhibited EGFR ubiquitination and maintained its protein stability by competing with the E3 ubiquitin ligase,c-Cbl.Additionally,EGFR was activated in liver cancer cells after sorafenib treatment,and SER-PINE2 knockdown-induced EGFR downregulation sig展开更多
Ischemic heart failure(HF)remains a leading cause of morbidity and mortality.Maintaining homeostasis of cardiac function and preventing cardiac remodeling deterioration are critical to halting HF progression.Methyltra...Ischemic heart failure(HF)remains a leading cause of morbidity and mortality.Maintaining homeostasis of cardiac function and preventing cardiac remodeling deterioration are critical to halting HF progression.Methyltransferase-like protein 13(Mettl13)has been shown to regulate protein translation efficiency by acting as a protein lysine methyltransferase,but its role in cardiac pathology remains unexplored.This study aims to characterize the roles and mechanisms of Mettl13 in cardiac contractile function and HF.We found that Mettl13 was downregulated in the failing hearts of mice post-myocardial infarction(MI)and in a cellular model of oxidative stress.Cardiomyocyte-specific overexpression of Mettl13 mediated by AAV9-Mettl13 attenuated cardiac contractile dysfunction and fibrosis in response to MI,while silencing of Mettl13 impaired cardiac function in normal mice.Moreover,Mettl13 overexpression abrogated the reduction in cell shortening,Ca^(2+)transient amplitude and SERCA2a protein levels in the cardiomyocytes of adult mice with MI.Conversely,knockdown of Mettl13 impaired the contractility of cardiomyocytes,and decreased Ca^(2+)transient amplitude and SERCA2a protein expression in vivo and in vitro.Mechanistically,Mettl13 impaired the stability of c-Cbl by inducing lysine methylation of c-Cbl,which in turn inhibited ubiquitination-dependent degradation of SERCA2a.Furthermore,the inhibitory effects of knocking down Mettl13 on SERCA2a protein expression and Ca^(2+)transients were partially rescued by silencing c-Cbl in H_(2)O_(2)-treated cardiomyocytes.In conclusion,our study uncovers a novel mechanism that involves the Mettl13/c-Cbl/SERCA2a axis in regulating cardiac contractile function and remodeling,and identifies Mettl13 as a novel therapeutic target for ischemic HF.展开更多
目的:研究c-Cbl和Cbl-b在胃癌组织中的表达及其临床意义.方法:选取2012-01/2014-12河南医学高等专科学校附属医院经手术切除得到了胃癌组织标本100例,设为观察组,另外选择其中20例癌旁正常胃组织标本设为对照组,将2组标本均制为组织芯片...目的:研究c-Cbl和Cbl-b在胃癌组织中的表达及其临床意义.方法:选取2012-01/2014-12河南医学高等专科学校附属医院经手术切除得到了胃癌组织标本100例,设为观察组,另外选择其中20例癌旁正常胃组织标本设为对照组,将2组标本均制为组织芯片,通过免疫组织化学方法对c-Cbl和Cbl-b表达的情况进行检测,研究其表达与临床病理特征间的关系.结果:c-Cbl和Cbl-b在观察组中的阳性率分别为72.0%与83.0%,均高于对照组(阳性率分别为20.0%与25.0%);c-Cbl在胃癌组织中的表达与胃癌的病理分期和浸润深度呈明显正相关,其表达量随着病理分期加重、浸润深度加深而增加.Cbl-b在胃癌组织中的表达与胃癌的病理分期和淋巴结是否转移呈明显正相关,其表达量随着病理分期加重、淋巴结发生转移而增加;c-Cbl和Cbl-b在胃癌组织中的表达呈正相关(P<0.05),两种蛋白同时呈强阳性表达者29例,病例的临床病理特征主要是分期晚、淋巴结转移数目多、肿瘤组织浸润程度深和分化程度差.结论:胃癌的发展与侵袭会增加c-C b l和Cbl-b表达的阳性率,c-Cbl和Cbl-b的检测对于胃癌的临床诊断有一定的临床意义与价值,值得推广.展开更多
Sef(similar expression to fgf genes)作为FGF信号通路中可诱导的拮抗分子相继在斑马鱼,小鼠,和人类中被鉴定出来,并进行了相应的功能研究.目前对于Sef蛋白本身稳定性的研究还未见报道.对c-Cbl对Sef稳定性的影响进行了研究.免疫荧光实...Sef(similar expression to fgf genes)作为FGF信号通路中可诱导的拮抗分子相继在斑马鱼,小鼠,和人类中被鉴定出来,并进行了相应的功能研究.目前对于Sef蛋白本身稳定性的研究还未见报道.对c-Cbl对Sef稳定性的影响进行了研究.免疫荧光实验表明Sef能够和c-Cbl蛋白在细胞中发生共定位,随后的免疫共沉淀实验证明Sef能够和c-Cbl发生相互作用.体内泛素化实验表明c-Cbl能够使Sef发生明显的泛素化作用.这种泛素化最终导致了Sef本身的剂量依赖性的降解.针对c-Cbl的siRNA表达也使Sef稳定细胞系的表达水平得到恢复.结果表明,c-Cbl对Sef的泛素化及降解可能作为一种调控拮抗因子的蛋白质水平从而最终调节信号通路的一种机制.展开更多
基金This work was supported by grants from the National Nat-ural Science Foundation of China(82070652 and 81870434)Department of Science and Technology of Zhejiang Province(2020C04003)+2 种基金the Chinese Academy of Medi-cal Sciences(019-I2M-5-030)the Jinan Microecological Biomedicine Shandong Laboratory(JNL-2022007B)the State Key Laboratory for Diagnosis and Treatment of Infectious Diseases(zz202302).
文摘Background:Liver cancer is a malignancy with high morbidity and mortality rates.Serpin family E member 2(SERPINE2)has been reported to play a key role in the metastasis of many tumors.In this study,we aimed to investigate the potential mechanism of SERPINE2 in liver cancer metastasis.Methods:The Cancer Genome Atlas database(TCGA),including DNA methy-lation and transcriptome sequencing data,was utilized to identify the crucial oncogene associated with DNA methylation and cancer progression in liver can-cer.Data from the TCGA and RNA sequencing for 94 pairs of liver cancer tissues were used to explore the correlation between SERPINE2 expression and clin-ical parameters of patients.DNA methylation sequencing was used to detect the DNA methylation levels in liver cancer tissues and cells.RNA sequencing,cytokine assays,immunoprecipitation(IP)and mass spectrometry(MS)assays,protein stability assays,and ubiquitination assays were performed to explore the regulatory mechanism of SERPINE2 in liver cancer metastasis.Patient-derived xenografts and tumor organoid models were established to determine the role of SERPINE2 in the treatment of liver cancer using sorafenib.Results:Based on the public database screening,SERPINE2 was identified as a tumor promoter regulated by DNA methylation.SERPINE2 expression was significantly higher in liver cancer tissues and was associated with the dismal prognosis in patients with liver cancer.SERPINE2 promoted liver cancer metas-tasis by enhancing cell pseudopodia formation,cell adhesion,cancer-associated fibroblast activation,extracellular matrix remodeling,and angiogenesis.IP/MS assays confirmed that SERPINE2 activated epidermal growth factor receptor(EGFR)and its downstream signaling pathways by interacting with EGFR.Mechanistically,SERPINE2 inhibited EGFR ubiquitination and maintained its protein stability by competing with the E3 ubiquitin ligase,c-Cbl.Additionally,EGFR was activated in liver cancer cells after sorafenib treatment,and SER-PINE2 knockdown-induced EGFR downregulation sig
基金supported by the National Natural Science Foundation of China (82273928,U21A20339)the Outstanding Youth Project of Natural Science Foundation of Heilongjiang Province (YQ2020H010)+2 种基金Youth Project of Scientific Research Institution of Heilongjiang Province (CZKYF2023-1-C047)CAMS Innovation Fund for Medical Sciences (CIFMS) (2019-I2M-5-078)Harbin Medical University Youth Talents Start-up Funding (2019-YQ-03)。
文摘Ischemic heart failure(HF)remains a leading cause of morbidity and mortality.Maintaining homeostasis of cardiac function and preventing cardiac remodeling deterioration are critical to halting HF progression.Methyltransferase-like protein 13(Mettl13)has been shown to regulate protein translation efficiency by acting as a protein lysine methyltransferase,but its role in cardiac pathology remains unexplored.This study aims to characterize the roles and mechanisms of Mettl13 in cardiac contractile function and HF.We found that Mettl13 was downregulated in the failing hearts of mice post-myocardial infarction(MI)and in a cellular model of oxidative stress.Cardiomyocyte-specific overexpression of Mettl13 mediated by AAV9-Mettl13 attenuated cardiac contractile dysfunction and fibrosis in response to MI,while silencing of Mettl13 impaired cardiac function in normal mice.Moreover,Mettl13 overexpression abrogated the reduction in cell shortening,Ca^(2+)transient amplitude and SERCA2a protein levels in the cardiomyocytes of adult mice with MI.Conversely,knockdown of Mettl13 impaired the contractility of cardiomyocytes,and decreased Ca^(2+)transient amplitude and SERCA2a protein expression in vivo and in vitro.Mechanistically,Mettl13 impaired the stability of c-Cbl by inducing lysine methylation of c-Cbl,which in turn inhibited ubiquitination-dependent degradation of SERCA2a.Furthermore,the inhibitory effects of knocking down Mettl13 on SERCA2a protein expression and Ca^(2+)transients were partially rescued by silencing c-Cbl in H_(2)O_(2)-treated cardiomyocytes.In conclusion,our study uncovers a novel mechanism that involves the Mettl13/c-Cbl/SERCA2a axis in regulating cardiac contractile function and remodeling,and identifies Mettl13 as a novel therapeutic target for ischemic HF.
文摘目的:研究c-Cbl和Cbl-b在胃癌组织中的表达及其临床意义.方法:选取2012-01/2014-12河南医学高等专科学校附属医院经手术切除得到了胃癌组织标本100例,设为观察组,另外选择其中20例癌旁正常胃组织标本设为对照组,将2组标本均制为组织芯片,通过免疫组织化学方法对c-Cbl和Cbl-b表达的情况进行检测,研究其表达与临床病理特征间的关系.结果:c-Cbl和Cbl-b在观察组中的阳性率分别为72.0%与83.0%,均高于对照组(阳性率分别为20.0%与25.0%);c-Cbl在胃癌组织中的表达与胃癌的病理分期和浸润深度呈明显正相关,其表达量随着病理分期加重、浸润深度加深而增加.Cbl-b在胃癌组织中的表达与胃癌的病理分期和淋巴结是否转移呈明显正相关,其表达量随着病理分期加重、淋巴结发生转移而增加;c-Cbl和Cbl-b在胃癌组织中的表达呈正相关(P<0.05),两种蛋白同时呈强阳性表达者29例,病例的临床病理特征主要是分期晚、淋巴结转移数目多、肿瘤组织浸润程度深和分化程度差.结论:胃癌的发展与侵袭会增加c-C b l和Cbl-b表达的阳性率,c-Cbl和Cbl-b的检测对于胃癌的临床诊断有一定的临床意义与价值,值得推广.
文摘Sef(similar expression to fgf genes)作为FGF信号通路中可诱导的拮抗分子相继在斑马鱼,小鼠,和人类中被鉴定出来,并进行了相应的功能研究.目前对于Sef蛋白本身稳定性的研究还未见报道.对c-Cbl对Sef稳定性的影响进行了研究.免疫荧光实验表明Sef能够和c-Cbl蛋白在细胞中发生共定位,随后的免疫共沉淀实验证明Sef能够和c-Cbl发生相互作用.体内泛素化实验表明c-Cbl能够使Sef发生明显的泛素化作用.这种泛素化最终导致了Sef本身的剂量依赖性的降解.针对c-Cbl的siRNA表达也使Sef稳定细胞系的表达水平得到恢复.结果表明,c-Cbl对Sef的泛素化及降解可能作为一种调控拮抗因子的蛋白质水平从而最终调节信号通路的一种机制.
