目的:寻找影响胃癌发生发展的衰老基因,构建内源竞争性RNA (ceRNA)调控网络,为发掘胃癌有效的诊断、预后生物标志物和治疗靶点提供依据。方法:从癌症基因组图谱(TCGA)数据库下载胃癌的表达数据和临床生存数据,在Aging Atlas数据库中获...目的:寻找影响胃癌发生发展的衰老基因,构建内源竞争性RNA (ceRNA)调控网络,为发掘胃癌有效的诊断、预后生物标志物和治疗靶点提供依据。方法:从癌症基因组图谱(TCGA)数据库下载胃癌的表达数据和临床生存数据,在Aging Atlas数据库中获取衰老基因,使用R软件中“DESeq2”程序包进行差异分析并采用Cox回归及Kaplan-Meier生存分析的方法筛选在胃癌中表达差异且与预后相关的衰老基因。通过Starbase数据库筛选具有靶向关系的miRNAs和lncRNAs,分别进行相关性分析和生存分析用于构建ceRNA调控网络。结果:对502个衰老基因分析显示SERPINE1基因在胃癌组织中显著高表达,且其高表达时患者预后较差。之后以SERPINE1作为关键基因构建一个与胃癌患者预后相关的ceRNA网络,其包含1个mRNA、1个miRNA和1个lncRNA,并以此基因构建胃癌的预后风险模型,最后进行通路、功能及免疫相关性分析。结论:本研究对衰老基因的分析及ceRNA网络的构建有助于进一步探索胃癌发生发展的分子机制,对发现预后标志物和治疗靶点至关重要。Objective: Searching for aging genes affecting the occurrence and development of gastric cancer and constructing competitive endogenous RNA (ceRNA) regulatory networks provide evidence for exploring effective diagnostic and prognostic biomarkers and therapeutic targets of gastric cancer. Methods: Expression data and clinical survival data of gastric cancer were downloaded from The Cancer Genome Atlas (TCGA) database. Senescence-related genes were obtained from the Aging Atlas database. Differential analysis was performed using the “DESeq2” package in R software. Cox regression and Kaplan-Meier survival analysis methods were employed to screen for senescence-related genes that are differentially expressed in gastric cancer and associated with prognosis. The Starbase database was used to identify miRNAs and lncRNAs with target relationships. Correlation analysis and survival analysis were conducted to construct the ceRNA regulatory network. Results: The analysis of 502 aging genes showed that the SERPINE1 gene was highly expressed in gastric cancer tissues, and the prognosis of patients was poor when it was highly expressed. Then SERPINE1 was used as the key gene to construct a ceRNA network correlated with survival in gastric cancer patients, which included one mRNA, one miRNA, and one lncRNA. The prognostic risk model of gastric cancer was constructed with the SERPINE1 gene. Finally, the pathway, functional enrichment analysis, and immune correlation analysis were carried out. Conclusion: The analysis of aging genes and the construction of the ceRNA network in this study are helpful to further explore the molecular mechanism of the occurrence and development of gastric cancer, which is crucial for the discovery of the prognostic markers and therapeutic targets.展开更多
BACKGROUND Gastric cancer(GC)is a highly lethal malignancy with a high incidence and mortality rate globally.Its development follows the Correa model,with intestinal metaplasia(IM)being a critical precursor to GC.Howe...BACKGROUND Gastric cancer(GC)is a highly lethal malignancy with a high incidence and mortality rate globally.Its development follows the Correa model,with intestinal metaplasia(IM)being a critical precursor to GC.However,the mechanisms underlying IM progression to GC remain unclear.This study explored ex-tracellular matrix(ECM)-related gene changes during IM progression to GC,aiming to identify biomarkers that could improve early diagnosis and treatment strategies for GC,ultimately enhancing patient outcomes.AIM To analyze transcriptome sequencing data,molecular biomarkers that can predict GC risk and monitor IM progression can be identified,providing new insights and strategies for preventing IM-GC transformation.METHODS Weighted gene co-expression network analysis served for confirming gene modules.Upregulated ECM-related genes were further tested using univariate Cox regression and least absolute shrinkage and selection operator analysis to select hub genes and construct a survival analysis model.The intestinal cell model was established by stimulating GES-1 cells with chenodeoxycholic acid.RESULTS Weighted gene co-expression network analysis identified 1709 differentially expressed genes from the GSE191275 dataset,while The Cancer Genome Atlas stomach adenocarcinoma revealed 4633 differentially expressed genes.The intersection of these datasets identified 71 upregulated and 171 downregulated genes,which were enriched in ECM-related pathways.Univariate Cox regression analysis identified six genes with prognostic significance,and least absolute shrinkage and selection operator regression pinpointed secreted protein acidic and rich in cysteine(SPARC)and SERPINE1 as non-zero coefficient genes.A prognostic model integrating clinical tumor node metastasis staging,age,SERPINE1,and SPARC was constructed.Immunohistochemistry analysis confirmed an increasing expression of SPARC protein from normal gastric mucosa(-),to IM(+-to+),and to GC(+to++),with significant differences(P<0.05).Western blot analysis demonstrated significantly higher SPARC expression in induced intestinal cells compared to GES-1.Furthermore,after SPARC knockdown in the human GC cell line HGC27,cell counting kit-8 and colony formation assays showed a reduction in cell proliferative ability,while the wound healing assay revealed impaired cell migration capacity.CONCLUSION Comprehensive analysis suggested that a model incorporating clinical tumor node metastasis staging,age,and SPARC/SERPINE1 expression served as a prognostic predictor for GC.Moreover,elevated SPARC expression in IM and GC suggests its potential as a proper biomarker to detect GC in early stage and as a novel therapeutic target,guiding clinical applications.展开更多
Background:Effective inhibition of pathological cardiac hypertrophy is critical for managing various cardiovascular diseases,especially in cold environments.The communication between cardiomyocytes and fibroblasts,med...Background:Effective inhibition of pathological cardiac hypertrophy is critical for managing various cardiovascular diseases,especially in cold environments.The communication between cardiomyocytes and fibroblasts,mediated by secreted proteins,plays a significant role in the development and progression of pathological cardiac hypertrophy.Serpin Family E Member 2(serpinE2),secreted by fibroblasts into the extracellular space,has been implicated in this process.However,whether serpinE2 can be internalized by cardiomyocytes and whether cold exposure influences this process remains unclear.Materials and methods:Mice were subjected to cold exposure(4°C,12 h/day for 8 weeks),and cardiac hypertrophy was induced by transverse aortic constriction(TAC).SerpinE2 expression was silenced by short interfering RNA(siRNA).Cardiac fibroblasts were stimulated with angiotensin II(Ang II)to induce serpinE2 secretion.Exogenous recombinant serpinE2,labeled with DyLight 488 or His-tag,was used to evaluate its internalization and functional role in cardiomyocytes.Internalization was inhibited by using antibodies against serpinE2,heparin,or endocytosis inhibitors(β-cyclodextrin,nystatin,dynasore,and chlorpromazine).Chromatin immunoprecipitation followed by quantitative polymerase chain reaction(PCR)was used to assess the binding of the transcription factor CDX1 to the serpinE2 promoter.Results:Cold exposure significantly increased serpinE2 mRNA and protein expression in mouse hearts.SerpinE2 levels were also upregulated in plasma and cardiac tissue following TAC.Knockdown of serpinE2 attenuated TAC-induced hypertrophy,restored left ventricular function,and reduced atrial natriuretic peptide,brain natriuretic peptide,andβ-myosin heavy chain fragment levels.Exogenous serpinE2 promoted cardiomyocyte hypertrophy,an effect that was reversed by serpinE2 knockdown.Co-culture with conditioned medium from Ang II-stimulated fibroblasts increased serpinE2 expression in cardiomyocytes.Exogenous serpinE2 was internalized via endocytosis,which was inhibited by antibodies,heparin,and endocytosis blockers.Internalized serpinE2 activated the protein kinase B(AKT)/β-catenin pathway in cardiomyocytes.CDX1 bound to the serpinE2 promoter and promoted its transcription in fibroblasts.CDX1 overexpression increased serpinE2 and collagen expression,while its suppression had the opposite effect.Administration of exogenous fibroblast growth factor 4(FGF4)or overexpression of FGF4 plasmid upregulated CDX1,serpinE2,and collagen expression in fibroblasts.Conclusions:SerpinE2 expression is responsive to cold stress and mediates intercellular communication between fibroblasts and cardiomyocytes.Fibroblast-secreted serpinE2 is internalized by cardiomyocytes via endocytosis,promoting hypertrophy through activation of the phosphatidylinositol 3-kinase(PI3K)-AKT/β-catenin pathway.The FGF4-CDX1 axis regulates serpinE2 expression and secretion in cardiac fibroblasts.展开更多
背景丝氨酸蛋白酶抑制剂家族E成员1(serine protease inhibitor family E member 1,SERPINE1)蛋白表达与多种癌细胞的增值、迁移和侵袭密切相关,目前缺乏对消化系统疾病的系统研究和报道.目的基于生物信息技术探究SERPINE1调节消化系统...背景丝氨酸蛋白酶抑制剂家族E成员1(serine protease inhibitor family E member 1,SERPINE1)蛋白表达与多种癌细胞的增值、迁移和侵袭密切相关,目前缺乏对消化系统疾病的系统研究和报道.目的基于生物信息技术探究SERPINE1调节消化系统癌症的内在机制.方法使用基因表达谱交互分析(gene expression profile analysis)进行SERPINE1表达分析,并使用Kaplan-Meier分析分析SERPINE1在临床预后中的潜力.结果SERPINE1表达与CD14、CD163以及CCL20相关,SERPINE1高表达可能通过促进巨噬细胞介导炎症,从而参与消化系统癌症的发生和发展.结论SERPINE1高表达可能通过促进巨噬细胞介导炎症,在消化系统癌症炎症和纤维化的调节中发挥重要作用.展开更多
BACKGROUND Colorectal cancer(CRC)is a major cause of cancer-related mortality,with limited therapeutic options for advanced stages.Simvastatin,primarily used to lower cholesterol,has shown potential as an anticancer a...BACKGROUND Colorectal cancer(CRC)is a major cause of cancer-related mortality,with limited therapeutic options for advanced stages.Simvastatin,primarily used to lower cholesterol,has shown potential as an anticancer agent.It may exert its effects by inhibiting SERPINE1,a protein implicated in CRC progression,and activating the cyclic guanosine monophosphate-protein kinase G(cGMP/PKG)signaling pathway.Given these findings,this study hypothesizes that simvastatin inhibits CRC cell proliferation and migration by downregulating SERPINE1 and activating the cGMP/PKG pathway,offering a novel therapeutic strategy for CRC management.AIM To study the effects of simvastatin on the function of colon cancer cells and to uncover the underlying mechanisms.METHODS NCM460,HCT-116,and SW620 cell lines were used for in vitro experiments with simvastatin at doses of 20μM,40μM,and 80μM.The Stitch database was used to analyze the target genes of simvastatin,whereas STRING was used to investigate SERPINE1 and its related pathways.HCT-116 and SW620 cells were transfected with single-cell RNA sequencing reveals SERPINE1 with or without Rp-8-Br-cGMP(a PKG inhibitor).Cell toxicity,proliferation,and migration were evaluated using the cell counting kit-8,colony formation,and Transwell assays,respectively.Apoptosis was analyzed via flow cytometry,and levels of reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH),and ferrous ion(Fe^(2+))were detected using commercial kits.Real-time polymerase chain reaction and western blotting were used to analyze gene expression.RESULTS Simvastatin dose-dependently inhibited the proliferation and migration of HCT-116 and SW620 cells while promoting apoptosis.It downregulated Ki-67,proliferating cell nuclear antigen,MMP2,and MMP9,and upregulated Bax,particularly at higher doses.Simvastatin increased the ROS,MDA,and Fe^(2+)levels while decreasing the GSH levels.It downregulated SLC7A11 and ferroportin and upregulated TRF1.SERPINE1 was identified as a core target,with related genes enriched in the cGMP/PKG pathway.SERPINE1 knockdown increased GUCY1B1 and PRKG1 levels,decreased cell viability,and altered oxidative stress markers,with the effects being reversed by Rp-8-Br-cGMP.CONCLUSION Simvastatin effectively inhibited the proliferation and migration of colon cancer cells and promoted apoptosis through the modulation of key targets,such as SERPINE1 and the cGMP/PKG signaling pathway.展开更多
BACKGROUND Gastric cancer(GC)is a malignant tumor originating from gastric mucosal epithelial cells that has high morbidity and mortality.microRNAs(miR)are important diagnostic markers and therapeutic targets in this ...BACKGROUND Gastric cancer(GC)is a malignant tumor originating from gastric mucosal epithelial cells that has high morbidity and mortality.microRNAs(miR)are important diagnostic markers and therapeutic targets in this disease.AIM To explore the mechanism of miR-125a-5p in the pathogenesis of GC.METHODS The expression levels of miR-125a-5p,SERPINE1 and DNMT1 in GC cells and tissues were detected by real-time polymerase chain reaction(PCR)and Western blotting.Methylation-specific PCR was used to detect the level of miR-125a-5p methylation.A cell counting kit 8 assay,scratch test,and a Transwell assay were performed to detect the proliferation,migration,and invasiveness of HGC27 cells,respectively.The expression of the epithelial mesenchymal transition(EMT)-related proteins E-cadherin,N-cadherin and vimentin in HGC27 cells was detected by Western blotting,while the expression of vimentin was detected by immunofluorescence.RESULTS This study revealed that miR-125a-5p was expressed at low levels in GC clinical samples and cells and that miR-125a-5p overexpression inhibited the proliferation,migration,invasiveness and EMT of GC cells.Mechanistically,miR-125a-5p can reduce GC cell proliferation,promote E-cadherin expression,inhibit N-cadherin and vimentin expression,and reduce the EMT of GC cells,thus constraining GC cells to a certain extent.Moreover,DNMT1 inhibited miR-125a-5p expression by increasing the methylation of the miR-125a-5p promoter,thereby promoting the expression of SERPINE1,which acts together with miR-125a-5p to exert antagonistic effects on GC.CONCLUSION Our study revealed that DNMT1 promoted SERPINE1 protein expression by inducing miR-125a-5p methylation,which led to the proliferation,migration and occurrence of EMT in GC cells.展开更多
BACKGROUND The upregulation of serpin family B member 5(SERPINB5)has been linked to the progression of rectal cancer.However,the specific roles and underlying mechanisms of SERPINB5 in rectal cancer are not fully unde...BACKGROUND The upregulation of serpin family B member 5(SERPINB5)has been linked to the progression of rectal cancer.However,the specific roles and underlying mechanisms of SERPINB5 in rectal cancer are not fully understood.AIM To investigate the roles and mechanisms of SERPINB5 in rectal cancer.METHODS SERPINB5 protein level in rectal cancer tissues and cell lines was measured through western blot analysis.SW480 cells were transfected with pcDNASERPINB5 or short-hairpin RNA targeting SERPINB5(sh-SERPINB5).Cell proliferation,invasion,and apoptosis were then evaluated.The interaction between SERPINB5 and heat shock protein 90 alpha class A member 1(HSP90AA1)was confirmed through a co-immunoprecipitation assay.Subsequently,pcDNAHSP90AA1 or sh-HSP90AA1 was transfected into SW480 cells,and cell progression was then detected.Moreover,rescue experiments were used to investigate the effect of the SERPINB5/HSP90AA1 axis on rectal cancer progression.Additionally,sh-SERPINB5-transfected SW480 cells were implanted into nude mice,and xenograft tumor growth was then evaluated.RESULTS SERPINB5 was prominently upregulated in rectal cancer tissues and cells.SERPINB5 overexpression increased SW480 cell proliferation and invasion while reducing apoptosis.In contrast,SERPINB5 knockdown had the opposite effects.Moreover,SERPINB5 could interact with HSP90AA1 and promote HSP90AA1 expression in SW480 cells.HSP90AA1 overexpression facilitated SW480 cell proliferation and invasion and restrained apoptosis.By contrast,HSP90AA1 knockdown suppressed cell progression.The upregulation of HSP90AA1 reversed the SERPINB5 silencing-mediated inhibition of SW480 cell progression.Additionally,SERPINB5 knockdown retarded the growth of rectal cancer tumors in vivo.CONCLUSION SERPINB5 knockdown inhibited rectal cancer cell proliferation and invasion and retarded xenograft tumor growth by inhibiting HSP90AA1 expression.展开更多
【目的】克隆牦牛丝氨酸蛋白酶抑制剂家族E成员1(serpin family E member 1,SERPINE1)基因序列,同时探究不同嫩度牦牛背最长肌组织中SERPINE1基因核苷酸序列差异,为进一步研究其对牦牛肉嫩度的影响提供试验数据。【方法】根据GenBank中...【目的】克隆牦牛丝氨酸蛋白酶抑制剂家族E成员1(serpin family E member 1,SERPINE1)基因序列,同时探究不同嫩度牦牛背最长肌组织中SERPINE1基因核苷酸序列差异,为进一步研究其对牦牛肉嫩度的影响提供试验数据。【方法】根据GenBank中牦牛SERPINE1基因序列设计特异性引物,通过PCR扩增克隆获得高、低嫩度牦牛背最长肌中的SERPINE1基因序列全长,并对其结构及编码蛋白进行生物信息学分析,通过实时荧光定量PCR检测不同嫩度牦牛背最长肌中SERPINE1基因表达量。【结果】高、低嫩度牦牛背最长肌组织中SERPINE1基因全长分别为8315和8318 bp,均编码402个氨基酸且氨基酸序列完全相同。高、低嫩度背最长肌组织中SERPINE1基因非编码序列中存在3个碱基缺失及22个碱基突变(4个碱基颠换、18个碱基转换)。牦牛SERPINE1基因核苷酸序列与普通牛、瘤牛、美洲野牛、绵羊、山羊、家猪、人、小鼠的相似性分别为99.26%、99.26%、99.59%、96.94%、97.02%、90.49%、86.52%和80.10%。SERPINE1蛋白是一个含有23个氨基酸信号肽的亲水性稳定膜外蛋白,位于细胞外,具有34个潜在磷酸化位点,包含1个反应中心环(reactive center loop,RCL)。SERPINE1蛋白二级结构主要由α-螺旋(44.78%)和无规则卷曲(35.32%)构成。实时荧光定量PCR结果显示,SERPINE1基因在高嫩度牦牛背最长肌中表达量极显著高于低嫩度牦牛背最长肌(P<0.01)。【结论】本研究成功从高、低嫩度牦牛背最长肌组织中克隆SERPINE1基因全长并进行了生物信息学特征分析,为后续研究SERPINE1基因参与调控牦牛肉嫩度机制提供理论参考。展开更多
BACKGROUND MicroRNAs(miRNAs)regulate gene expression and play a critical role in cancer physiology.However,there is still a limited understanding of the function and regulatory mechanism of miRNAs in gastric cancer(GC...BACKGROUND MicroRNAs(miRNAs)regulate gene expression and play a critical role in cancer physiology.However,there is still a limited understanding of the function and regulatory mechanism of miRNAs in gastric cancer(GC).AIM To investigate the role and molecular mechanism of miRNA-145-5p(miR145-5p)in the progression of GC.METHODS Real-time polymerase chain reaction(RT-PCR)was used to detect miRNA expression in human GC tissues and cells.The ability of cancer cells to migrate and invade was assessed using wound-healing and transwell assays,respectively.Cell proliferation was measured using cell counting kit-8 and colony formation assays,and apoptosis was evaluated using flow cytometry.Expression of the epithelial-mesenchymal transition(EMT)-associated protein was determined by Western blot.Targets of miR-145-5p were predicated using bioinformatics analysis and verified using a dual-luciferase reporter system.Serpin family E member 1(SERPINE1)expression in GC tissues and cells was evaluated using RT-PCR and immunohistochemical staining.The correlation between SERPINE1 expression and overall patient survival was determined using Kaplan-Meier plot analysis.The association between SERPINE1 and GC progression was also tested.A rescue experiment of SERPINE1 overexpression was conducted to verify the relationship between this protein and miR-145-5p.The mechanism by which miR-145-5p influences GC progression was further explored by assessing tumor formation in nude mice.RESULTS GC tissues and cells had reduced miR-145-5p expression and SERPINE1 was identified as a direct target of this miRNA.Overexpression of miR-145-5p was associated with decreased GC cell proliferation,invasion,migration,and EMT,and these effects were reversed by forcing SERPINE1 expression.Kaplan-Meier plot analysis revealed that patients with higher SERPINE1 expression had a shorter survival rate than those with lower SERPINE1 expression.Nude mouse tumorigenesis experiments confirmed that miR-145-5p targets SERPINE1 to regulate extracellular signal-regulated kinase-1/2(ERK1/2).CONCLUSION This study found that miR-145-5p inhibits tumor progression and is expressed in lower amounts in patients with GC.MiR-145-5p was found to affect GC cell proliferation,migration,and invasion by negatively regulating SERPINE1 levels and controlling the ERK1/2 pathway.展开更多
BACKGROUND Serpin peptidase inhibitor clade H member 1(SERPINH1)was initially recognized as an oncogene implicated in various human malignancies.Nevertheless,the clinical relevance and functional implications of SERPI...BACKGROUND Serpin peptidase inhibitor clade H member 1(SERPINH1)was initially recognized as an oncogene implicated in various human malignancies.Nevertheless,the clinical relevance and functional implications of SERPINH1 in colorectal cancer(CRC)remain largely elusive.AIM To investigate the effects of SERPINH1 on CRC cells and its specific mechanism.METHODS Quantitative real-time polymerase chain reaction,western blotting analysis,The Cancer Genome Atlas data mining and immunohistochemistry were employed to examine SERPINH1 expression in CRC cell lines and tissues.A series of in-vitro assays were performed to demonstrate the function of SERPINH1 and its possible mechanisms in CRC.RESULTS SERPINH1 demonstrated elevated expression levels in both CRC cells and tissues,manifested at both mRNA and protein tiers.Elevated SERPINH1 levels correlated closely with advanced T stage,lymph node involvement,and distant metastasis,exhibiting a significant association with poorer overall survival among CRC patients.Subsequent investigations unveiled that SERPINH1 overexpression notably bolstered CRC cell proliferation,invasion,and migration in vitro,while conversely,SERPINH1 knockdown elicited the opposite effects.Gene set enrichment analysis underscored a correlation between SERPINH1 upregulation and genes associated with cell cycle regulation.Our findings underscored the capacity of heightened SERPINH1 levels to expedite G1/S phase cell cycle progression via phosphatidylinositol 3-kinase/AKT/mechanistic target of rapamycin pathway activation,thereby facilitating CRC cell invasion and migration.CONCLUSION These findings imply a crucial involvement of SERPINH1 in the advancement and escalation of CRC,potentially positioning it as a novel candidate for prognostic assessment and therapeutic intervention in CRC management.展开更多
人体的抗凝系统主要有抗凝血酶（antithrombin，AT）系统（包括抗凝血酶和肝素，主要抑制具有丝氨酸蛋白酶活性的凝血因子，如：FXa，凝血酶等）和蛋白 C 系统（包括蛋白 C，蛋白 S，血栓调节蛋白，内皮细胞蛋白 C 受体等，主要灭活凝...人体的抗凝系统主要有抗凝血酶（antithrombin，AT）系统（包括抗凝血酶和肝素，主要抑制具有丝氨酸蛋白酶活性的凝血因子，如：FXa，凝血酶等）和蛋白 C 系统（包括蛋白 C，蛋白 S，血栓调节蛋白，内皮细胞蛋白 C 受体等，主要灭活凝血因子Ⅴ和Ⅷ等），而抗凝血酶系统中的抗凝血酶是人体最重要的抗凝因子之一，约占抗凝活性的70％。抗凝血酶是一种丝氨酸蛋白酶（serpins）抑制剂，主要具有抑制丝氨酸蛋白酶活性的凝血酶、FXa 等因子，在人体抗凝系统中发挥着重要作用。展开更多
文摘目的:寻找影响胃癌发生发展的衰老基因,构建内源竞争性RNA (ceRNA)调控网络,为发掘胃癌有效的诊断、预后生物标志物和治疗靶点提供依据。方法:从癌症基因组图谱(TCGA)数据库下载胃癌的表达数据和临床生存数据,在Aging Atlas数据库中获取衰老基因,使用R软件中“DESeq2”程序包进行差异分析并采用Cox回归及Kaplan-Meier生存分析的方法筛选在胃癌中表达差异且与预后相关的衰老基因。通过Starbase数据库筛选具有靶向关系的miRNAs和lncRNAs,分别进行相关性分析和生存分析用于构建ceRNA调控网络。结果:对502个衰老基因分析显示SERPINE1基因在胃癌组织中显著高表达,且其高表达时患者预后较差。之后以SERPINE1作为关键基因构建一个与胃癌患者预后相关的ceRNA网络,其包含1个mRNA、1个miRNA和1个lncRNA,并以此基因构建胃癌的预后风险模型,最后进行通路、功能及免疫相关性分析。结论:本研究对衰老基因的分析及ceRNA网络的构建有助于进一步探索胃癌发生发展的分子机制,对发现预后标志物和治疗靶点至关重要。Objective: Searching for aging genes affecting the occurrence and development of gastric cancer and constructing competitive endogenous RNA (ceRNA) regulatory networks provide evidence for exploring effective diagnostic and prognostic biomarkers and therapeutic targets of gastric cancer. Methods: Expression data and clinical survival data of gastric cancer were downloaded from The Cancer Genome Atlas (TCGA) database. Senescence-related genes were obtained from the Aging Atlas database. Differential analysis was performed using the “DESeq2” package in R software. Cox regression and Kaplan-Meier survival analysis methods were employed to screen for senescence-related genes that are differentially expressed in gastric cancer and associated with prognosis. The Starbase database was used to identify miRNAs and lncRNAs with target relationships. Correlation analysis and survival analysis were conducted to construct the ceRNA regulatory network. Results: The analysis of 502 aging genes showed that the SERPINE1 gene was highly expressed in gastric cancer tissues, and the prognosis of patients was poor when it was highly expressed. Then SERPINE1 was used as the key gene to construct a ceRNA network correlated with survival in gastric cancer patients, which included one mRNA, one miRNA, and one lncRNA. The prognostic risk model of gastric cancer was constructed with the SERPINE1 gene. Finally, the pathway, functional enrichment analysis, and immune correlation analysis were carried out. Conclusion: The analysis of aging genes and the construction of the ceRNA network in this study are helpful to further explore the molecular mechanism of the occurrence and development of gastric cancer, which is crucial for the discovery of the prognostic markers and therapeutic targets.
基金Supported by the Inner Mongolia Autonomous Region Health Science and Technology Program,No.202201441Qingmiao Plan of Baotou Medical College,No.BYJJ-ZRQM202230。
文摘BACKGROUND Gastric cancer(GC)is a highly lethal malignancy with a high incidence and mortality rate globally.Its development follows the Correa model,with intestinal metaplasia(IM)being a critical precursor to GC.However,the mechanisms underlying IM progression to GC remain unclear.This study explored ex-tracellular matrix(ECM)-related gene changes during IM progression to GC,aiming to identify biomarkers that could improve early diagnosis and treatment strategies for GC,ultimately enhancing patient outcomes.AIM To analyze transcriptome sequencing data,molecular biomarkers that can predict GC risk and monitor IM progression can be identified,providing new insights and strategies for preventing IM-GC transformation.METHODS Weighted gene co-expression network analysis served for confirming gene modules.Upregulated ECM-related genes were further tested using univariate Cox regression and least absolute shrinkage and selection operator analysis to select hub genes and construct a survival analysis model.The intestinal cell model was established by stimulating GES-1 cells with chenodeoxycholic acid.RESULTS Weighted gene co-expression network analysis identified 1709 differentially expressed genes from the GSE191275 dataset,while The Cancer Genome Atlas stomach adenocarcinoma revealed 4633 differentially expressed genes.The intersection of these datasets identified 71 upregulated and 171 downregulated genes,which were enriched in ECM-related pathways.Univariate Cox regression analysis identified six genes with prognostic significance,and least absolute shrinkage and selection operator regression pinpointed secreted protein acidic and rich in cysteine(SPARC)and SERPINE1 as non-zero coefficient genes.A prognostic model integrating clinical tumor node metastasis staging,age,SERPINE1,and SPARC was constructed.Immunohistochemistry analysis confirmed an increasing expression of SPARC protein from normal gastric mucosa(-),to IM(+-to+),and to GC(+to++),with significant differences(P<0.05).Western blot analysis demonstrated significantly higher SPARC expression in induced intestinal cells compared to GES-1.Furthermore,after SPARC knockdown in the human GC cell line HGC27,cell counting kit-8 and colony formation assays showed a reduction in cell proliferative ability,while the wound healing assay revealed impaired cell migration capacity.CONCLUSION Comprehensive analysis suggested that a model incorporating clinical tumor node metastasis staging,age,and SPARC/SERPINE1 expression served as a prognostic predictor for GC.Moreover,elevated SPARC expression in IM and GC suggests its potential as a proper biomarker to detect GC in early stage and as a novel therapeutic target,guiding clinical applications.
基金supported by the National Natural Science Foundation of China(82370279,82170299,82330011,82003757).
文摘Background:Effective inhibition of pathological cardiac hypertrophy is critical for managing various cardiovascular diseases,especially in cold environments.The communication between cardiomyocytes and fibroblasts,mediated by secreted proteins,plays a significant role in the development and progression of pathological cardiac hypertrophy.Serpin Family E Member 2(serpinE2),secreted by fibroblasts into the extracellular space,has been implicated in this process.However,whether serpinE2 can be internalized by cardiomyocytes and whether cold exposure influences this process remains unclear.Materials and methods:Mice were subjected to cold exposure(4°C,12 h/day for 8 weeks),and cardiac hypertrophy was induced by transverse aortic constriction(TAC).SerpinE2 expression was silenced by short interfering RNA(siRNA).Cardiac fibroblasts were stimulated with angiotensin II(Ang II)to induce serpinE2 secretion.Exogenous recombinant serpinE2,labeled with DyLight 488 or His-tag,was used to evaluate its internalization and functional role in cardiomyocytes.Internalization was inhibited by using antibodies against serpinE2,heparin,or endocytosis inhibitors(β-cyclodextrin,nystatin,dynasore,and chlorpromazine).Chromatin immunoprecipitation followed by quantitative polymerase chain reaction(PCR)was used to assess the binding of the transcription factor CDX1 to the serpinE2 promoter.Results:Cold exposure significantly increased serpinE2 mRNA and protein expression in mouse hearts.SerpinE2 levels were also upregulated in plasma and cardiac tissue following TAC.Knockdown of serpinE2 attenuated TAC-induced hypertrophy,restored left ventricular function,and reduced atrial natriuretic peptide,brain natriuretic peptide,andβ-myosin heavy chain fragment levels.Exogenous serpinE2 promoted cardiomyocyte hypertrophy,an effect that was reversed by serpinE2 knockdown.Co-culture with conditioned medium from Ang II-stimulated fibroblasts increased serpinE2 expression in cardiomyocytes.Exogenous serpinE2 was internalized via endocytosis,which was inhibited by antibodies,heparin,and endocytosis blockers.Internalized serpinE2 activated the protein kinase B(AKT)/β-catenin pathway in cardiomyocytes.CDX1 bound to the serpinE2 promoter and promoted its transcription in fibroblasts.CDX1 overexpression increased serpinE2 and collagen expression,while its suppression had the opposite effect.Administration of exogenous fibroblast growth factor 4(FGF4)or overexpression of FGF4 plasmid upregulated CDX1,serpinE2,and collagen expression in fibroblasts.Conclusions:SerpinE2 expression is responsive to cold stress and mediates intercellular communication between fibroblasts and cardiomyocytes.Fibroblast-secreted serpinE2 is internalized by cardiomyocytes via endocytosis,promoting hypertrophy through activation of the phosphatidylinositol 3-kinase(PI3K)-AKT/β-catenin pathway.The FGF4-CDX1 axis regulates serpinE2 expression and secretion in cardiac fibroblasts.
文摘背景丝氨酸蛋白酶抑制剂家族E成员1(serine protease inhibitor family E member 1,SERPINE1)蛋白表达与多种癌细胞的增值、迁移和侵袭密切相关,目前缺乏对消化系统疾病的系统研究和报道.目的基于生物信息技术探究SERPINE1调节消化系统癌症的内在机制.方法使用基因表达谱交互分析(gene expression profile analysis)进行SERPINE1表达分析,并使用Kaplan-Meier分析分析SERPINE1在临床预后中的潜力.结果SERPINE1表达与CD14、CD163以及CCL20相关,SERPINE1高表达可能通过促进巨噬细胞介导炎症,从而参与消化系统癌症的发生和发展.结论SERPINE1高表达可能通过促进巨噬细胞介导炎症,在消化系统癌症炎症和纤维化的调节中发挥重要作用.
基金Supported by the Jiangsu Province Key Research and Development Plan(Social Development)Project,No.BE2021611。
文摘BACKGROUND Colorectal cancer(CRC)is a major cause of cancer-related mortality,with limited therapeutic options for advanced stages.Simvastatin,primarily used to lower cholesterol,has shown potential as an anticancer agent.It may exert its effects by inhibiting SERPINE1,a protein implicated in CRC progression,and activating the cyclic guanosine monophosphate-protein kinase G(cGMP/PKG)signaling pathway.Given these findings,this study hypothesizes that simvastatin inhibits CRC cell proliferation and migration by downregulating SERPINE1 and activating the cGMP/PKG pathway,offering a novel therapeutic strategy for CRC management.AIM To study the effects of simvastatin on the function of colon cancer cells and to uncover the underlying mechanisms.METHODS NCM460,HCT-116,and SW620 cell lines were used for in vitro experiments with simvastatin at doses of 20μM,40μM,and 80μM.The Stitch database was used to analyze the target genes of simvastatin,whereas STRING was used to investigate SERPINE1 and its related pathways.HCT-116 and SW620 cells were transfected with single-cell RNA sequencing reveals SERPINE1 with or without Rp-8-Br-cGMP(a PKG inhibitor).Cell toxicity,proliferation,and migration were evaluated using the cell counting kit-8,colony formation,and Transwell assays,respectively.Apoptosis was analyzed via flow cytometry,and levels of reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH),and ferrous ion(Fe^(2+))were detected using commercial kits.Real-time polymerase chain reaction and western blotting were used to analyze gene expression.RESULTS Simvastatin dose-dependently inhibited the proliferation and migration of HCT-116 and SW620 cells while promoting apoptosis.It downregulated Ki-67,proliferating cell nuclear antigen,MMP2,and MMP9,and upregulated Bax,particularly at higher doses.Simvastatin increased the ROS,MDA,and Fe^(2+)levels while decreasing the GSH levels.It downregulated SLC7A11 and ferroportin and upregulated TRF1.SERPINE1 was identified as a core target,with related genes enriched in the cGMP/PKG pathway.SERPINE1 knockdown increased GUCY1B1 and PRKG1 levels,decreased cell viability,and altered oxidative stress markers,with the effects being reversed by Rp-8-Br-cGMP.CONCLUSION Simvastatin effectively inhibited the proliferation and migration of colon cancer cells and promoted apoptosis through the modulation of key targets,such as SERPINE1 and the cGMP/PKG signaling pathway.
基金the Research Program of the Science and Technology Department of Yunnan Province,No.202101AY070001-204.
文摘BACKGROUND Gastric cancer(GC)is a malignant tumor originating from gastric mucosal epithelial cells that has high morbidity and mortality.microRNAs(miR)are important diagnostic markers and therapeutic targets in this disease.AIM To explore the mechanism of miR-125a-5p in the pathogenesis of GC.METHODS The expression levels of miR-125a-5p,SERPINE1 and DNMT1 in GC cells and tissues were detected by real-time polymerase chain reaction(PCR)and Western blotting.Methylation-specific PCR was used to detect the level of miR-125a-5p methylation.A cell counting kit 8 assay,scratch test,and a Transwell assay were performed to detect the proliferation,migration,and invasiveness of HGC27 cells,respectively.The expression of the epithelial mesenchymal transition(EMT)-related proteins E-cadherin,N-cadherin and vimentin in HGC27 cells was detected by Western blotting,while the expression of vimentin was detected by immunofluorescence.RESULTS This study revealed that miR-125a-5p was expressed at low levels in GC clinical samples and cells and that miR-125a-5p overexpression inhibited the proliferation,migration,invasiveness and EMT of GC cells.Mechanistically,miR-125a-5p can reduce GC cell proliferation,promote E-cadherin expression,inhibit N-cadherin and vimentin expression,and reduce the EMT of GC cells,thus constraining GC cells to a certain extent.Moreover,DNMT1 inhibited miR-125a-5p expression by increasing the methylation of the miR-125a-5p promoter,thereby promoting the expression of SERPINE1,which acts together with miR-125a-5p to exert antagonistic effects on GC.CONCLUSION Our study revealed that DNMT1 promoted SERPINE1 protein expression by inducing miR-125a-5p methylation,which led to the proliferation,migration and occurrence of EMT in GC cells.
基金Supported by the Medical Science Research Project Plan of the Hebei Provincial Health Commission,No.20210027.
文摘BACKGROUND The upregulation of serpin family B member 5(SERPINB5)has been linked to the progression of rectal cancer.However,the specific roles and underlying mechanisms of SERPINB5 in rectal cancer are not fully understood.AIM To investigate the roles and mechanisms of SERPINB5 in rectal cancer.METHODS SERPINB5 protein level in rectal cancer tissues and cell lines was measured through western blot analysis.SW480 cells were transfected with pcDNASERPINB5 or short-hairpin RNA targeting SERPINB5(sh-SERPINB5).Cell proliferation,invasion,and apoptosis were then evaluated.The interaction between SERPINB5 and heat shock protein 90 alpha class A member 1(HSP90AA1)was confirmed through a co-immunoprecipitation assay.Subsequently,pcDNAHSP90AA1 or sh-HSP90AA1 was transfected into SW480 cells,and cell progression was then detected.Moreover,rescue experiments were used to investigate the effect of the SERPINB5/HSP90AA1 axis on rectal cancer progression.Additionally,sh-SERPINB5-transfected SW480 cells were implanted into nude mice,and xenograft tumor growth was then evaluated.RESULTS SERPINB5 was prominently upregulated in rectal cancer tissues and cells.SERPINB5 overexpression increased SW480 cell proliferation and invasion while reducing apoptosis.In contrast,SERPINB5 knockdown had the opposite effects.Moreover,SERPINB5 could interact with HSP90AA1 and promote HSP90AA1 expression in SW480 cells.HSP90AA1 overexpression facilitated SW480 cell proliferation and invasion and restrained apoptosis.By contrast,HSP90AA1 knockdown suppressed cell progression.The upregulation of HSP90AA1 reversed the SERPINB5 silencing-mediated inhibition of SW480 cell progression.Additionally,SERPINB5 knockdown retarded the growth of rectal cancer tumors in vivo.CONCLUSION SERPINB5 knockdown inhibited rectal cancer cell proliferation and invasion and retarded xenograft tumor growth by inhibiting HSP90AA1 expression.
文摘【目的】克隆牦牛丝氨酸蛋白酶抑制剂家族E成员1(serpin family E member 1,SERPINE1)基因序列,同时探究不同嫩度牦牛背最长肌组织中SERPINE1基因核苷酸序列差异,为进一步研究其对牦牛肉嫩度的影响提供试验数据。【方法】根据GenBank中牦牛SERPINE1基因序列设计特异性引物,通过PCR扩增克隆获得高、低嫩度牦牛背最长肌中的SERPINE1基因序列全长,并对其结构及编码蛋白进行生物信息学分析,通过实时荧光定量PCR检测不同嫩度牦牛背最长肌中SERPINE1基因表达量。【结果】高、低嫩度牦牛背最长肌组织中SERPINE1基因全长分别为8315和8318 bp,均编码402个氨基酸且氨基酸序列完全相同。高、低嫩度背最长肌组织中SERPINE1基因非编码序列中存在3个碱基缺失及22个碱基突变(4个碱基颠换、18个碱基转换)。牦牛SERPINE1基因核苷酸序列与普通牛、瘤牛、美洲野牛、绵羊、山羊、家猪、人、小鼠的相似性分别为99.26%、99.26%、99.59%、96.94%、97.02%、90.49%、86.52%和80.10%。SERPINE1蛋白是一个含有23个氨基酸信号肽的亲水性稳定膜外蛋白,位于细胞外,具有34个潜在磷酸化位点,包含1个反应中心环(reactive center loop,RCL)。SERPINE1蛋白二级结构主要由α-螺旋(44.78%)和无规则卷曲(35.32%)构成。实时荧光定量PCR结果显示,SERPINE1基因在高嫩度牦牛背最长肌中表达量极显著高于低嫩度牦牛背最长肌(P<0.01)。【结论】本研究成功从高、低嫩度牦牛背最长肌组织中克隆SERPINE1基因全长并进行了生物信息学特征分析,为后续研究SERPINE1基因参与调控牦牛肉嫩度机制提供理论参考。
文摘BACKGROUND MicroRNAs(miRNAs)regulate gene expression and play a critical role in cancer physiology.However,there is still a limited understanding of the function and regulatory mechanism of miRNAs in gastric cancer(GC).AIM To investigate the role and molecular mechanism of miRNA-145-5p(miR145-5p)in the progression of GC.METHODS Real-time polymerase chain reaction(RT-PCR)was used to detect miRNA expression in human GC tissues and cells.The ability of cancer cells to migrate and invade was assessed using wound-healing and transwell assays,respectively.Cell proliferation was measured using cell counting kit-8 and colony formation assays,and apoptosis was evaluated using flow cytometry.Expression of the epithelial-mesenchymal transition(EMT)-associated protein was determined by Western blot.Targets of miR-145-5p were predicated using bioinformatics analysis and verified using a dual-luciferase reporter system.Serpin family E member 1(SERPINE1)expression in GC tissues and cells was evaluated using RT-PCR and immunohistochemical staining.The correlation between SERPINE1 expression and overall patient survival was determined using Kaplan-Meier plot analysis.The association between SERPINE1 and GC progression was also tested.A rescue experiment of SERPINE1 overexpression was conducted to verify the relationship between this protein and miR-145-5p.The mechanism by which miR-145-5p influences GC progression was further explored by assessing tumor formation in nude mice.RESULTS GC tissues and cells had reduced miR-145-5p expression and SERPINE1 was identified as a direct target of this miRNA.Overexpression of miR-145-5p was associated with decreased GC cell proliferation,invasion,migration,and EMT,and these effects were reversed by forcing SERPINE1 expression.Kaplan-Meier plot analysis revealed that patients with higher SERPINE1 expression had a shorter survival rate than those with lower SERPINE1 expression.Nude mouse tumorigenesis experiments confirmed that miR-145-5p targets SERPINE1 to regulate extracellular signal-regulated kinase-1/2(ERK1/2).CONCLUSION This study found that miR-145-5p inhibits tumor progression and is expressed in lower amounts in patients with GC.MiR-145-5p was found to affect GC cell proliferation,migration,and invasion by negatively regulating SERPINE1 levels and controlling the ERK1/2 pathway.
基金Supported by Ruian Natural Science Foundation,No.MS2021008.
文摘BACKGROUND Serpin peptidase inhibitor clade H member 1(SERPINH1)was initially recognized as an oncogene implicated in various human malignancies.Nevertheless,the clinical relevance and functional implications of SERPINH1 in colorectal cancer(CRC)remain largely elusive.AIM To investigate the effects of SERPINH1 on CRC cells and its specific mechanism.METHODS Quantitative real-time polymerase chain reaction,western blotting analysis,The Cancer Genome Atlas data mining and immunohistochemistry were employed to examine SERPINH1 expression in CRC cell lines and tissues.A series of in-vitro assays were performed to demonstrate the function of SERPINH1 and its possible mechanisms in CRC.RESULTS SERPINH1 demonstrated elevated expression levels in both CRC cells and tissues,manifested at both mRNA and protein tiers.Elevated SERPINH1 levels correlated closely with advanced T stage,lymph node involvement,and distant metastasis,exhibiting a significant association with poorer overall survival among CRC patients.Subsequent investigations unveiled that SERPINH1 overexpression notably bolstered CRC cell proliferation,invasion,and migration in vitro,while conversely,SERPINH1 knockdown elicited the opposite effects.Gene set enrichment analysis underscored a correlation between SERPINH1 upregulation and genes associated with cell cycle regulation.Our findings underscored the capacity of heightened SERPINH1 levels to expedite G1/S phase cell cycle progression via phosphatidylinositol 3-kinase/AKT/mechanistic target of rapamycin pathway activation,thereby facilitating CRC cell invasion and migration.CONCLUSION These findings imply a crucial involvement of SERPINH1 in the advancement and escalation of CRC,potentially positioning it as a novel candidate for prognostic assessment and therapeutic intervention in CRC management.
文摘人体的抗凝系统主要有抗凝血酶（antithrombin，AT）系统（包括抗凝血酶和肝素，主要抑制具有丝氨酸蛋白酶活性的凝血因子，如：FXa，凝血酶等）和蛋白 C 系统（包括蛋白 C，蛋白 S，血栓调节蛋白，内皮细胞蛋白 C 受体等，主要灭活凝血因子Ⅴ和Ⅷ等），而抗凝血酶系统中的抗凝血酶是人体最重要的抗凝因子之一，约占抗凝活性的70％。抗凝血酶是一种丝氨酸蛋白酶（serpins）抑制剂，主要具有抑制丝氨酸蛋白酶活性的凝血酶、FXa 等因子，在人体抗凝系统中发挥着重要作用。
