目的探讨细胞色素P450酶2W1(cytochrome P450 family 2 subfamily W member 1,CYP2W1)、微小RNA-491-5p(microRNA-491-5p,miR-491-5p)在肝细胞癌(hepatocellular cancer,HCC)组织中的表达与临床病理特征的关系和预后的相关性分析。方法...目的探讨细胞色素P450酶2W1(cytochrome P450 family 2 subfamily W member 1,CYP2W1)、微小RNA-491-5p(microRNA-491-5p,miR-491-5p)在肝细胞癌(hepatocellular cancer,HCC)组织中的表达与临床病理特征的关系和预后的相关性分析。方法选取2019年2月~2021年2月在我院进行手术治疗的120例HCC作为研究对象,手术过程中取HCC组织和癌旁组织标本。实时荧光定量PCR(qRT-PCR)法检测mRNA表达水平,免疫组化法检测蛋白表达水平。分析HCC组织和癌旁组织中miR-491-5p与CYP2W1 mRNA表达的差异及二者表达与患者临床病理特征的相关性,并做Kaplan-Meier生存分析曲线。结果HCC组织中CYP2W1 mRNA表达水平显著高于癌旁组织,miR-491-5p表达水平显著低于癌旁组织(P<0.05);HCC组织中CYP2W1蛋白阳性表达率显著高于癌旁组织(χ^(2)=23.721,P<0.001)。HCC组织中miR-491-5p与CYP2W1 mRNA表达水平呈负相关(r=-0.417,P<0.001)。CYP2W1、miR-491-5p表达与HCC患者TNM分期、肿瘤分化程度、淋巴结转移和病理分级存在显著相关性(P<0.05)。Kaplan-Meier生存分析曲线显示,CYP2W1阳性表达组3年累积生存率为31.4%,低于CYP2W1阴性表达组的63.8%(log-rankχ^(2)=12.243,P=0.000);miR-491-5p高表达组3年累积生存率为61.3%,显著高于miR-491-5p低表达组的37.9%(log-rankχ^(2)=4.490,P=0.034)。复发组HCC组织中CYP2W1 mRNA表达水平显著高于未复发组,miR-491-5p表达水平显著低于未复发组(P<0.05);转移组HCC组织中CYP2W1 mRNA表达水平显著高于未转移组,miR-491-5p表达水平显著低于未转移组(P<0.05)。结论HCC组织中CYP2W1高表达,miR-491-5p低表达,二者与HCC患者的部分临床病理特征和预后存在显著相关性。展开更多
BACKGROUND Shifting from the inflammatory to the proliferative phase represents a pivotal step during managing diabetic foot ulcers(DFUs);however,existing medical interventions remain insufficient.MicroRNAs(miRs)highl...BACKGROUND Shifting from the inflammatory to the proliferative phase represents a pivotal step during managing diabetic foot ulcers(DFUs);however,existing medical interventions remain insufficient.MicroRNAs(miRs)highlight notable capacity for accelerating the repair process of DFUs.Previous research has demonstrated which miR-122-5p regulates matrix metalloproteinases under diabetic conditions,thereby influencing extracellular matrix dynamics.AIM To investigate the impact of miR-122-5p on the transition from the inflammatory to the proliferative stage in DFU.METHODS Analysis for miR-122-5p expression in skin tissues from diabetic ulcer patients and mice was analyzed using quantitative real-time polymerase chain reaction(qRT-PCR).A diabetic wound healing model induced by streptozotocin was used,with mice receiving intradermal injections of adeno-associated virus-DJ encoding empty vector or miR-122.Skin tissues were retrieved at 3,7,and 14 days after injury for gene expression analysis,histology,immunohistochemistry,and network studies.The study explored miR-122-5p’s role in macrophage-fibroblast interactions and its effect on transitioning from inflammation to proliferation in DFU healing.RESULTS High-throughput sequencing revealed miR-122-5p as crucial for DFU healing.qRT-PCR showed significant upregulation of miR-122-5p within diabetic skin among DFU individuals and mice.Western blot,along with immunohistochemical and enzyme-linked immunosorbent assay,demonstrating the upregulation of inflammatory mediators(hypoxia inducible factor-1α,matrix metalloproteinase 9,tumor necrosis factor-α)and reduced fibrosis markers(fibronectin 1,α-smooth muscle actin)by targeting vascular endothelial growth factor.Fluorescence in situ hybridization indicated its expression localized to epidermal keratinocytes and fibroblasts in diabetic mice.Immunofluorescence revealed enhanced increased presence of M1 macrophages and reduced M2 polarization,highlighting its role in inflammation.MiR-122-5p elevated inflammatory cytokine levels while suppressing fibrotic activity from fibroblasts exposed to macrophage-derived media,highlighting its pivotal role in regulating DFU healing.CONCLUSION MiR-122-5p impedes cutaneous healing of diabetic mice via enhancing inflammation and inhibiting fibrosis,offering insights into miR roles in human skin wound repair.展开更多
Elemene is widely recognized as an effective anti-cancer compound and is routinely administered in Chinese clinical settings for the management of several solid tumors,including non-small cell lung cancer(NSCLC).Howev...Elemene is widely recognized as an effective anti-cancer compound and is routinely administered in Chinese clinical settings for the management of several solid tumors,including non-small cell lung cancer(NSCLC).However,its detailed molecular mechanism has not been adequately demonstrated.In this research,it was demonstrated that elemene effectively curtailed NSCLC growth in the patient-derived xenograft(PDX)model.Mechanistically,employing high-throughput screening techniques and subsequent biochemical validations such as microscale thermophoresis(MST),microRNA-145-5p(miR-145-5p)was pinpointed as a critical target through which elemene exerts its anti-tumor effects.Interestingly,elemene serves as a binding stabilizer for miR-145-5p,demonstrating a strong binding affinity(dissociation constant(KD)=0.39±0.17μg/mL)and preventing its degradation both in vitro and in vivo,while not interfering with the synthesis of the primary microRNA transcripts(pri-miRNAs)and precursor miRNAs(pre-miRNAs).The stabilization of miR-145-5p by elemene resulted in an increased level of this miRNA,subsequently suppressing NSCLC progression through the miR-145-5p/mitogen-activated protein kinase kinase kinase 3(MAP3K3)/nuclear factor kappaB(NF-κB)pathway.Our findings provide a new perspective on revealing the interaction patterns between clinical anti-tumor drugs and miRNAs.展开更多
文摘目的探讨细胞色素P450酶2W1(cytochrome P450 family 2 subfamily W member 1,CYP2W1)、微小RNA-491-5p(microRNA-491-5p,miR-491-5p)在肝细胞癌(hepatocellular cancer,HCC)组织中的表达与临床病理特征的关系和预后的相关性分析。方法选取2019年2月~2021年2月在我院进行手术治疗的120例HCC作为研究对象,手术过程中取HCC组织和癌旁组织标本。实时荧光定量PCR(qRT-PCR)法检测mRNA表达水平,免疫组化法检测蛋白表达水平。分析HCC组织和癌旁组织中miR-491-5p与CYP2W1 mRNA表达的差异及二者表达与患者临床病理特征的相关性,并做Kaplan-Meier生存分析曲线。结果HCC组织中CYP2W1 mRNA表达水平显著高于癌旁组织,miR-491-5p表达水平显著低于癌旁组织(P<0.05);HCC组织中CYP2W1蛋白阳性表达率显著高于癌旁组织(χ^(2)=23.721,P<0.001)。HCC组织中miR-491-5p与CYP2W1 mRNA表达水平呈负相关(r=-0.417,P<0.001)。CYP2W1、miR-491-5p表达与HCC患者TNM分期、肿瘤分化程度、淋巴结转移和病理分级存在显著相关性(P<0.05)。Kaplan-Meier生存分析曲线显示,CYP2W1阳性表达组3年累积生存率为31.4%,低于CYP2W1阴性表达组的63.8%(log-rankχ^(2)=12.243,P=0.000);miR-491-5p高表达组3年累积生存率为61.3%,显著高于miR-491-5p低表达组的37.9%(log-rankχ^(2)=4.490,P=0.034)。复发组HCC组织中CYP2W1 mRNA表达水平显著高于未复发组,miR-491-5p表达水平显著低于未复发组(P<0.05);转移组HCC组织中CYP2W1 mRNA表达水平显著高于未转移组,miR-491-5p表达水平显著低于未转移组(P<0.05)。结论HCC组织中CYP2W1高表达,miR-491-5p低表达,二者与HCC患者的部分临床病理特征和预后存在显著相关性。
基金Supported by the National Natural Science Foundation of China,No.82274528.
文摘BACKGROUND Shifting from the inflammatory to the proliferative phase represents a pivotal step during managing diabetic foot ulcers(DFUs);however,existing medical interventions remain insufficient.MicroRNAs(miRs)highlight notable capacity for accelerating the repair process of DFUs.Previous research has demonstrated which miR-122-5p regulates matrix metalloproteinases under diabetic conditions,thereby influencing extracellular matrix dynamics.AIM To investigate the impact of miR-122-5p on the transition from the inflammatory to the proliferative stage in DFU.METHODS Analysis for miR-122-5p expression in skin tissues from diabetic ulcer patients and mice was analyzed using quantitative real-time polymerase chain reaction(qRT-PCR).A diabetic wound healing model induced by streptozotocin was used,with mice receiving intradermal injections of adeno-associated virus-DJ encoding empty vector or miR-122.Skin tissues were retrieved at 3,7,and 14 days after injury for gene expression analysis,histology,immunohistochemistry,and network studies.The study explored miR-122-5p’s role in macrophage-fibroblast interactions and its effect on transitioning from inflammation to proliferation in DFU healing.RESULTS High-throughput sequencing revealed miR-122-5p as crucial for DFU healing.qRT-PCR showed significant upregulation of miR-122-5p within diabetic skin among DFU individuals and mice.Western blot,along with immunohistochemical and enzyme-linked immunosorbent assay,demonstrating the upregulation of inflammatory mediators(hypoxia inducible factor-1α,matrix metalloproteinase 9,tumor necrosis factor-α)and reduced fibrosis markers(fibronectin 1,α-smooth muscle actin)by targeting vascular endothelial growth factor.Fluorescence in situ hybridization indicated its expression localized to epidermal keratinocytes and fibroblasts in diabetic mice.Immunofluorescence revealed enhanced increased presence of M1 macrophages and reduced M2 polarization,highlighting its role in inflammation.MiR-122-5p elevated inflammatory cytokine levels while suppressing fibrotic activity from fibroblasts exposed to macrophage-derived media,highlighting its pivotal role in regulating DFU healing.CONCLUSION MiR-122-5p impedes cutaneous healing of diabetic mice via enhancing inflammation and inhibiting fibrosis,offering insights into miR roles in human skin wound repair.
基金supported by the National Natural Science Foundation of China(Grant No.:82225048)the Dalian Science and Technology Leading Talents Project,China(Grant No.:2019RD15)Sanming Project of Medicine in Shenzhen,China(Grant No.:SZZYSM202106004).
文摘Elemene is widely recognized as an effective anti-cancer compound and is routinely administered in Chinese clinical settings for the management of several solid tumors,including non-small cell lung cancer(NSCLC).However,its detailed molecular mechanism has not been adequately demonstrated.In this research,it was demonstrated that elemene effectively curtailed NSCLC growth in the patient-derived xenograft(PDX)model.Mechanistically,employing high-throughput screening techniques and subsequent biochemical validations such as microscale thermophoresis(MST),microRNA-145-5p(miR-145-5p)was pinpointed as a critical target through which elemene exerts its anti-tumor effects.Interestingly,elemene serves as a binding stabilizer for miR-145-5p,demonstrating a strong binding affinity(dissociation constant(KD)=0.39±0.17μg/mL)and preventing its degradation both in vitro and in vivo,while not interfering with the synthesis of the primary microRNA transcripts(pri-miRNAs)and precursor miRNAs(pre-miRNAs).The stabilization of miR-145-5p by elemene resulted in an increased level of this miRNA,subsequently suppressing NSCLC progression through the miR-145-5p/mitogen-activated protein kinase kinase kinase 3(MAP3K3)/nuclear factor kappaB(NF-κB)pathway.Our findings provide a new perspective on revealing the interaction patterns between clinical anti-tumor drugs and miRNAs.
