Objective:To investigate the effect of microRNA(miR-184)on regulating the genesis.development and prolifration of glioma cells.Methods:Lipidosome was used to transfect miR-184 mimic and inhibitor to glioma cell ling,a...Objective:To investigate the effect of microRNA(miR-184)on regulating the genesis.development and prolifration of glioma cells.Methods:Lipidosome was used to transfect miR-184 mimic and inhibitor to glioma cell ling,and the cell proliferation ability changes were determined by MTT and plate cloning experiment after the transfection.WB test was used to measure the levels of cyclinD1,p27 and FOXO3.Meanwhile.QPCR was used to detect miR-184expression in glioma cell line.glioma tissues and adjacent tissues.Luciferase experiment was used to test 3'UTR gene targeting regulation of miR-184 and FOXO3.Results:QPCR results showed a significaat lower miR-184 expression level in glioma cell line and glioma tissues than that in juxtacancerous tissue.MTT and plate cloning experiments have shown that after overexpressing of miR-184,the cell proliferation capacity of glioma U87 and T98G was signifieantly increased,which was significantly inhibited after the inhibition of miN-184.WB results showed a lower expression level of p27 in U87 and T98G cells,and a higher expression level of cyclind1after over-expressing of miR-184 wss observed.However.a lower expression level of eyelinD1and a higher expression level of p27 after the inhibition of miR-184.The luciferase activity was inhibited after the over-expressing of miR-184.Conclusions:MiR-184 can affect the proliferation abilities of glioma cells and regulate the cell cycle related protein.It plays an important role in the occurrence and development of gliomas.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 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supported by National Science Foundation(Grant No.81372721)
文摘Objective:To investigate the effect of microRNA(miR-184)on regulating the genesis.development and prolifration of glioma cells.Methods:Lipidosome was used to transfect miR-184 mimic and inhibitor to glioma cell ling,and the cell proliferation ability changes were determined by MTT and plate cloning experiment after the transfection.WB test was used to measure the levels of cyclinD1,p27 and FOXO3.Meanwhile.QPCR was used to detect miR-184expression in glioma cell line.glioma tissues and adjacent tissues.Luciferase experiment was used to test 3'UTR gene targeting regulation of miR-184 and FOXO3.Results:QPCR results showed a significaat lower miR-184 expression level in glioma cell line and glioma tissues than that in juxtacancerous tissue.MTT and plate cloning experiments have shown that after overexpressing of miR-184,the cell proliferation capacity of glioma U87 and T98G was signifieantly increased,which was significantly inhibited after the inhibition of miN-184.WB results showed a lower expression level of p27 in U87 and T98G cells,and a higher expression level of cyclind1after over-expressing of miR-184 wss observed.However.a lower expression level of eyelinD1and a higher expression level of p27 after the inhibition of miR-184.The luciferase activity was inhibited after the over-expressing of miR-184.Conclusions:MiR-184 can affect the proliferation abilities of glioma cells and regulate the cell cycle related protein.It plays an important role in the occurrence and development of gliomas.
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.
