Background:MicroRNAs(miRNAs)play critical roles in corneal development and functional homeostasis.Our previous study identified miR-184 as one of the most highly expressed miRNAs in the corneal epithelium.Even though ...Background:MicroRNAs(miRNAs)play critical roles in corneal development and functional homeostasis.Our previous study identified miR-184 as one of the most highly expressed miRNAs in the corneal epithelium.Even though its expression level plummeted dramatically during corneal epithelial wound healing(CEWH),its precise role in mediating corneal epithelial renewal was unresolved.The present study aimed to reveal the function and mechanism of miR-184 in regulating CEWH.Methods:Quantitative RT-PCR analysis characterized the miR-184 expression pattern during CEWH in mice.Ectopic miR-184 injection determined its effect on this process in vivo.We evaluated the effects of miR-184 and its target genes on the proliferation,cell cycle,and migration of human corneal epithelial cells(HCECs)using MTS,flow cytometry,and wound-healing assay,respectively.Bioinformatic analysis,in conjunction with gene microarray analysis and cell-based luciferase assays,pinpointed gene targets of miR-184 contributing to CEWH.Results:MiR-184 underwent marked downregulation during mouse CEWH.Ectopic miR-184 overexpression delayed this process in mice.Furthermore,miR-184 transfection into HCECs significantly inhibited cell proliferation,cell cycle progression,and cell migration.MiR-184 directly targeted CDC25A,CARM1,and LASP1,and downregulated their expression in HCECs.CARM1 downregulation inhibited both HCEC proliferation and migration,whereas a decrease in LASP1 gene expression only inhibited migration.Conclusions:Our results demonstrate that miR-184 inhibits corneal epithelial cell proliferation and migration via targeting CDC25A,CARM1,and LASP1,suggesting it acts as a negative modulator during CEWH.Therefore,identifying strategies to suppress miR-184 expression levels has the potential to promote CEWH.展开更多
Background:MicroRNAs(miRNAs)play critical roles in corneal development and functional homeostasis.Our previous study identified miR-184 as one of the most highly expressed miRNAs in the corneal epithelium.Even though ...Background:MicroRNAs(miRNAs)play critical roles in corneal development and functional homeostasis.Our previous study identified miR-184 as one of the most highly expressed miRNAs in the corneal epithelium.Even though its expression level plummeted dramatically during corneal epithelial wound healing(CEWH),its precise role in mediating corneal epithelial renewal was unresolved.The present study aimed to reveal the function and mechanism of miR-184 in regulating CEWH.Methods:Quantitative reverse transcriptase polymerase chain reaction(RT-PCR)analysis characterized the miR-184 expression pattern during CEWH in mice.Ectopic miR-184 injection determined its effect on this process in vivo.We evaluated the effects of miR-184 and its target genes on the proliferation,cell cycle,and migration of human corneal epithelial cells(HCECs)using MTS,flow cytometry,and wound healing assay,respectively.Bioinformatic analysis,in conjunction with gene microarray analysis and cell-based luciferase assays,pinpointed gene targets of miR-184 contributing to CEWH.Results:MiR-184 underwent marked downregulation during mouse CEWH.Ectopic miR-184 overexpression delayed this process in mice.Furthermore,miR-184 transfection into HCECs significantly inhibited cell proliferation,cell cycle progression,and cell migration.MiR-184 directly targeted CDC25A,CARMI,and LASP1,and downregulated their expression in HCECs.CARM1 downregulation inhibited both HCEC proliferation and migration,whereas a decrease in LASPI gene expression only inhibited migration.Conclusi ons:Our results dem on strate that miR-184 inhibits corneal epithelial cell proliferation and migration via targeting CDC25A,CARMI,and LASPI,suggesting it acts as a negative modulator during CEWH.Therefore,identifying strategies to suppress miR-184 expression levels has the potential to promote CEWH.展开更多
基金This work was supported,in part,by the 973 Project(2012CB722303)from the Ministry of Science and Technology of China,Science Foundation of Wenzhou Medical University(QTJ11020)the Science and Technology Project of Wenzhou(Grant No.Y20160188).
文摘Background:MicroRNAs(miRNAs)play critical roles in corneal development and functional homeostasis.Our previous study identified miR-184 as one of the most highly expressed miRNAs in the corneal epithelium.Even though its expression level plummeted dramatically during corneal epithelial wound healing(CEWH),its precise role in mediating corneal epithelial renewal was unresolved.The present study aimed to reveal the function and mechanism of miR-184 in regulating CEWH.Methods:Quantitative RT-PCR analysis characterized the miR-184 expression pattern during CEWH in mice.Ectopic miR-184 injection determined its effect on this process in vivo.We evaluated the effects of miR-184 and its target genes on the proliferation,cell cycle,and migration of human corneal epithelial cells(HCECs)using MTS,flow cytometry,and wound-healing assay,respectively.Bioinformatic analysis,in conjunction with gene microarray analysis and cell-based luciferase assays,pinpointed gene targets of miR-184 contributing to CEWH.Results:MiR-184 underwent marked downregulation during mouse CEWH.Ectopic miR-184 overexpression delayed this process in mice.Furthermore,miR-184 transfection into HCECs significantly inhibited cell proliferation,cell cycle progression,and cell migration.MiR-184 directly targeted CDC25A,CARM1,and LASP1,and downregulated their expression in HCECs.CARM1 downregulation inhibited both HCEC proliferation and migration,whereas a decrease in LASP1 gene expression only inhibited migration.Conclusions:Our results demonstrate that miR-184 inhibits corneal epithelial cell proliferation and migration via targeting CDC25A,CARM1,and LASP1,suggesting it acts as a negative modulator during CEWH.Therefore,identifying strategies to suppress miR-184 expression levels has the potential to promote CEWH.
基金supported,in part,by the 973 Project(Grant No.2012CB722303)the Ministry of Science and Technology of China,Science Foundation of Wenzhou Medical University(Grant No.QTJ11020)the Science and Technology Project of Wenzhou(Grant No.Y20160188).
文摘Background:MicroRNAs(miRNAs)play critical roles in corneal development and functional homeostasis.Our previous study identified miR-184 as one of the most highly expressed miRNAs in the corneal epithelium.Even though its expression level plummeted dramatically during corneal epithelial wound healing(CEWH),its precise role in mediating corneal epithelial renewal was unresolved.The present study aimed to reveal the function and mechanism of miR-184 in regulating CEWH.Methods:Quantitative reverse transcriptase polymerase chain reaction(RT-PCR)analysis characterized the miR-184 expression pattern during CEWH in mice.Ectopic miR-184 injection determined its effect on this process in vivo.We evaluated the effects of miR-184 and its target genes on the proliferation,cell cycle,and migration of human corneal epithelial cells(HCECs)using MTS,flow cytometry,and wound healing assay,respectively.Bioinformatic analysis,in conjunction with gene microarray analysis and cell-based luciferase assays,pinpointed gene targets of miR-184 contributing to CEWH.Results:MiR-184 underwent marked downregulation during mouse CEWH.Ectopic miR-184 overexpression delayed this process in mice.Furthermore,miR-184 transfection into HCECs significantly inhibited cell proliferation,cell cycle progression,and cell migration.MiR-184 directly targeted CDC25A,CARMI,and LASP1,and downregulated their expression in HCECs.CARM1 downregulation inhibited both HCEC proliferation and migration,whereas a decrease in LASPI gene expression only inhibited migration.Conclusi ons:Our results dem on strate that miR-184 inhibits corneal epithelial cell proliferation and migration via targeting CDC25A,CARMI,and LASPI,suggesting it acts as a negative modulator during CEWH.Therefore,identifying strategies to suppress miR-184 expression levels has the potential to promote CEWH.
