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
Background:Hepatocellular carcinoma(HCC)typically begins inconspicuously and progresses swiftly,leading to most patients being diagnosed at an advanced stage.Accordingly,a pressing priority is to clarify the developme...Background:Hepatocellular carcinoma(HCC)typically begins inconspicuously and progresses swiftly,leading to most patients being diagnosed at an advanced stage.Accordingly,a pressing priority is to clarify the development mechanisms of HCC and devise efficient intervention and treatment protocols.Methods:An upstream miRNA of solute carrier transporter family 1 member 5(SLC1A5)was predicted to bemiR-122-5p by various databases,and a dual-luciferase reporter gene assay was used to verify the SLC1A5-and miR-122-5p-targeting relationship.SLC1A5 and miR-122-5p expression in HCC cells was quantitatively assessed using quantitative reverse transcription polymerase chain reaction(qRT–PCR).Western blotting was used to evaluate SLC1A5 expression in HCC cells.To determine the effects of the ferroptosis inducers erastin and L-g-glutamyl-p-nitroanilide(GPNA)on Huh-7 and HepG2 cell viability,a Cell Counting Kit-8 assay was performed.Additionally,various assay kits were used to assess malondialdehyde(MDA),Fe^(2+),and reactive oxygen species(ROS)levels inHCC cells.Moreover,anHCC tumor model was established in nude mice to investigate the growth of HCC tumors overexpressing miR-122-5p.Results:miR-122-5p downregulated SLC1A5 levels.MiR-122-5p knockdown inhibited erastin-promoted ferroptosis,whereas miR-122-5p overexpression promoted ferroptosis.After knocking down miR-122-5p,the MDA,Fe^(2+),and ROS levels in Huh-7 and HepG2 cells decreased,whereas overexpressing miR-122-5p increased the MDA,Fe^(2+),and ROS levels.Following the addition of GPNA,the cells experienced decreased viability and increased MDA levels,which in turn inhibited ferroptosis.Knockdown of SLC1A5 partially reversed the ferroptosis-inhibiting effect induced by knocking downmiR-122-5p.Additionally,the overexpression of miR-122-5p hindered HCC development in vivo.Conclusion:miR-122-5p downregulates SLC1A5,which promotes lipid peroxidation and iron accumulation and inhibits glutamine transport,ultimately causing ferroptosis in HCC cells.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
Objective Glioblastoma(GBM)is the most common,invasive,and malignant primary brain tumor with a poor prognosis and high recurrence rate.It’s known that some microRNAs(miRNAs)which are associated with tumorigenesis an...Objective Glioblastoma(GBM)is the most common,invasive,and malignant primary brain tumor with a poor prognosis and high recurrence rate.It’s known that some microRNAs(miRNAs)which are associated with tumorigenesis and progression can be considered as prognostic and therapeutic targets in tumors including GBM.This study aims to highlight the potential role of the core miRNAs in GBM and their potential use as a prognostic and therapeutic biomarker.Methods Differentially expressed miRNAs(DEmiRNAs)were identified in GBM by integrating miRNA-sequencing results and a GBM microarray dataset from the Gene Expression Omnibus(GEO)database through bioinformatics tools.The dysregulated miRNAs were identified by survival analysis through Chinese Glioma Genome Atlas(CGGA).Target genes of the dysregulated miRNAs were predicted on MiRWalk and miRTarBase database.TAM2.0 database,Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways analysis were used to analyze the function of the dysregulated miRNAs.Subsequently,protein-protein interaction(PPI)network analysis was used to identify the top 20 hub targets of the up-regulated and down-regulated miRNAs,respectively.Then,core miRNAs in GBM were identified by constructing dysregulated miRNA-differentially expressed hub gene networks.Validation of the core miRNAs expression was detected in 41 GBM tissues compared to 8 normal brain tissues.Furthermore,the potential biomarkers were identified by clinical correlation analysis and survival analysis.Results Totally,68 intersecting DEmiRNAs were identified,40 of which were upregulated and the other 28 miRNAs were downregulated.Two upregulated and 4 downregulated miRNAs showed prognostic significance.Most differentially expressed hub genes were regulated by the miR-28-5p and miR-1224-5p,which were respectively upregulated and downregulated in GBM.The correlation between miR-1224-5p level and recurrence was statistically significant(P=0.011).Survival analysis showed that high miR-28-5p level and high miR-1224-5p level were both associated with better prognosis.Moreover,high miR-1224-5p level was an independent prognosis factor for GBM patients according to the cox regression analysis.Conclusion MiRNA-1224-5p could be a potential target for the prognosis and treatment in GBM.展开更多
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Hubei Provincial Key Research and Development Program(2023BCB126)Hubei University of ChineseMedicine“14th Five-Year Plan”Outstanding Discipline TeamConstruction Project(HBUCM[2022]No.90).
文摘Background:Hepatocellular carcinoma(HCC)typically begins inconspicuously and progresses swiftly,leading to most patients being diagnosed at an advanced stage.Accordingly,a pressing priority is to clarify the development mechanisms of HCC and devise efficient intervention and treatment protocols.Methods:An upstream miRNA of solute carrier transporter family 1 member 5(SLC1A5)was predicted to bemiR-122-5p by various databases,and a dual-luciferase reporter gene assay was used to verify the SLC1A5-and miR-122-5p-targeting relationship.SLC1A5 and miR-122-5p expression in HCC cells was quantitatively assessed using quantitative reverse transcription polymerase chain reaction(qRT–PCR).Western blotting was used to evaluate SLC1A5 expression in HCC cells.To determine the effects of the ferroptosis inducers erastin and L-g-glutamyl-p-nitroanilide(GPNA)on Huh-7 and HepG2 cell viability,a Cell Counting Kit-8 assay was performed.Additionally,various assay kits were used to assess malondialdehyde(MDA),Fe^(2+),and reactive oxygen species(ROS)levels inHCC cells.Moreover,anHCC tumor model was established in nude mice to investigate the growth of HCC tumors overexpressing miR-122-5p.Results:miR-122-5p downregulated SLC1A5 levels.MiR-122-5p knockdown inhibited erastin-promoted ferroptosis,whereas miR-122-5p overexpression promoted ferroptosis.After knocking down miR-122-5p,the MDA,Fe^(2+),and ROS levels in Huh-7 and HepG2 cells decreased,whereas overexpressing miR-122-5p increased the MDA,Fe^(2+),and ROS levels.Following the addition of GPNA,the cells experienced decreased viability and increased MDA levels,which in turn inhibited ferroptosis.Knockdown of SLC1A5 partially reversed the ferroptosis-inhibiting effect induced by knocking downmiR-122-5p.Additionally,the overexpression of miR-122-5p hindered HCC development in vivo.Conclusion:miR-122-5p downregulates SLC1A5,which promotes lipid peroxidation and iron accumulation and inhibits glutamine transport,ultimately causing ferroptosis in HCC cells.
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.
基金the National Natural Science Foundation of China(No.81960453 and No.81860445)the Natural Science Foundation of Guangxi Province(No.2018GXNSFAA050151 and No.2018GXNSFAA281251)+2 种基金the Basic Ability Improvement Project for Young and Middle-aged Teachers in Colleges and Universities of Guangxi(No.2020KY03039)the Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor(Guangxi Medical University)Ministry of Education(No.GK2018-09,No.GKE 2019-08,and No.GKE-ZZ202006).
文摘Objective Glioblastoma(GBM)is the most common,invasive,and malignant primary brain tumor with a poor prognosis and high recurrence rate.It’s known that some microRNAs(miRNAs)which are associated with tumorigenesis and progression can be considered as prognostic and therapeutic targets in tumors including GBM.This study aims to highlight the potential role of the core miRNAs in GBM and their potential use as a prognostic and therapeutic biomarker.Methods Differentially expressed miRNAs(DEmiRNAs)were identified in GBM by integrating miRNA-sequencing results and a GBM microarray dataset from the Gene Expression Omnibus(GEO)database through bioinformatics tools.The dysregulated miRNAs were identified by survival analysis through Chinese Glioma Genome Atlas(CGGA).Target genes of the dysregulated miRNAs were predicted on MiRWalk and miRTarBase database.TAM2.0 database,Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways analysis were used to analyze the function of the dysregulated miRNAs.Subsequently,protein-protein interaction(PPI)network analysis was used to identify the top 20 hub targets of the up-regulated and down-regulated miRNAs,respectively.Then,core miRNAs in GBM were identified by constructing dysregulated miRNA-differentially expressed hub gene networks.Validation of the core miRNAs expression was detected in 41 GBM tissues compared to 8 normal brain tissues.Furthermore,the potential biomarkers were identified by clinical correlation analysis and survival analysis.Results Totally,68 intersecting DEmiRNAs were identified,40 of which were upregulated and the other 28 miRNAs were downregulated.Two upregulated and 4 downregulated miRNAs showed prognostic significance.Most differentially expressed hub genes were regulated by the miR-28-5p and miR-1224-5p,which were respectively upregulated and downregulated in GBM.The correlation between miR-1224-5p level and recurrence was statistically significant(P=0.011).Survival analysis showed that high miR-28-5p level and high miR-1224-5p level were both associated with better prognosis.Moreover,high miR-1224-5p level was an independent prognosis factor for GBM patients according to the cox regression analysis.Conclusion MiRNA-1224-5p could be a potential target for the prognosis and treatment in GBM.
