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Inhibition of miR‐205 impairs the wound‐healing process in human corneal epithelial cells by stimulating Kir4.1(KCNJ10)
Author(s) -
Lin Daohong,
Halivolic Adna,
Bellner Lars,
yue peng
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.lb682
Subject(s) - wound healing , transfection , western blot , microrna , microbiology and biotechnology , small hairpin rna , endogeny , cell culture , biology , chemistry , immunology , gene knockdown , gene , endocrinology , biochemistry , genetics
The aim of this study is to explore the role of miR‐205, the second most abundant microRNA expressed in human cornea epithelial cells (HCE) in modulating healing process of the cell following the scratch (wound). The scratch of HCE cells increased significantly the expression of miR‐205. Moreover, inhibition of miR‐205 with antagomer (pZip‐miR‐205) significantly impairs the healing process of HCE cells. However, inhibition of endogenous K channels with Ba 2+ partially restored the rate of healing process in HCE cells transfected with pZip‐miR‐205, suggesting that the inhibition of HCE healing process by miR‐205 antagomer was partially the result of stimulation of K channel activity. The patch‐clamp experiments further demonstrated that expression of miR‐205 antagomer increased the endogenous K currents in HCE cells in comparison to the cells transfected with vector alone. Analyzing the seed sequence of miR‐205 reveals that 3′UTR of KCNJ10 contains potential complementary matching sequence of miR‐205. Moreover, Western blot analysis demonstrated expression of pZip‐miR‐205 increased the protein expression of KCNJ10 but not KCNJ16 in HCE. Also, Western blot showed that expression of KCNJ10 was decreased 24 hrs in the post‐wound HCE cells. Finally, the role of KCNJ 10 in modulating the healing process of HCEC was further demonstrated by experiments in which down‐regulation of the K channel with kcnj10‐shRNA mimic the effect of Ba 2+ and partially restored the healing process of HCE cells transfected with miR‐205 antagomer. We conclude that miR‐205 plays an important role in facilitating the wound‐healing process in HCE, an effect was partially achieved by inhibiting the inwardly‐rectifying K channel.