Antagomir dependent microRNA-205 reduction enhances adhesion ability of human corneal epithelial keratinocytes.
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 microRNA-205 increased the number of phosphorylated FAK and phosphorylated Pax, and decreased filamentous actin.


CONCLUSIONS
Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.
Li
Jun
J
Department of Biochemistry, Institute of Basic Medical Sciences, Chinese Academyof Medical Sciences & Peking Union Medical College, Beijing 100005, China.
Bai
Hua
H
Zhu
Yong
Y
Wang
Xiao-yan
XY
Wang
Fang
F
Zhang
Jun-wu
JW
Lavker
Robert M
RM
Yu
Jia
J
D001483:Base Sequence; D002448:Cell Adhesion / Q000235:genetics*; D002478:Cells, Cultured; D019573:Epithelium, Corneal / Q000166:cytology*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D035683:MicroRNAs / Q000037:antagonists & inhibitors*; D015345:Oligonucleotide Probes
D035683:MicroRNAs; D015345:Oligonucleotide Probes