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Objective To investigate the effect of microRNA-205 reduction by antagomirs on adhesion ability of normal human corneal epithelial keratinocytes(NHCEKs).Methods Antagomir-205,complementary and inhibitory to microRNA-205,was used to suppress endogenous microRNA-205 in NHCEKs.The adhesion ability of treated NHCEKs was then assessed by cell adhesion assay.Immunoblot and immunohistochemistry were conducted to determine the level of two focal adhesion-related proteins,focal adhesion kinase(FAK) and paxillin(Pax).Phalloidin staining was performed to measure the level of filamentous actin in antagomir-treated NHCEKs.Results Antagomir-205 markedly reduced the level of microRNA-205 in NHCEKs and significantly enhanced adhesion ability of NHCEKs(P<0.01).Further protein analysis validated that inhibition of mi-croRNA-205 increased the number of phosphorylated FAK and phosphorylated Pax,and decreased filamen-tous actin.Conclusion Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.
Objective To investigate the effect of microRNA-205 reduction by antagomirs on adhesion ability of normal human corneal epithelial keratinocytes (NHCEKs). Methods Antagomir-205, complementary and inhibitory to microRNA-205, was used to suppress endogenous microRNA- 205 in NHCEKs.The adhesion ability of treated NHCEKs was then assessed by cell adhesion assay. Immunoblot and immunohistochemistry were conducted to determine the level of two focal adhesion-related proteins, focal adhesion kinase (FAK) and paxillin (Pax). Phalloidin staining was performed to measure the level The results of Antagomir-205 markedly reduced the level of microRNA-205 in NHCEKs and significantly enhanced adhesion ability of NHCEKs (P <0.01). Further protein analysis validated that inhibition of mi-croRNA-205 increased the number of phosphorylated FAK and phosphorylated Pax, and decreased filamen-tous actin. Confluence Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.