Antagomir Dependent MicroRNA-205 Reduction Enhances Adhesion Ability of Human Corneal Epithelial Keratinocytes

doi:10.1016/S1001-9294(10)60024-7

Chinese Medical Sciences Journal ›› 2010, Vol. 25 ›› Issue (2): 65-70.doi: 10.1016/S1001-9294(10)60024-7

• Original Article • 下一篇

Antagomir Dependent MicroRNA-205 Reduction Enhances Adhesion Ability of Human Corneal Epithelial Keratinocytes

Jun Li^1,2†, Hua Bai^3†, Yong Zhu¹, Xiao-yan Wang¹, Fang Wang¹, Jun-wu Zhang¹, Robert M. Lavker⁴, and Jia Yu^1*

^a Department of Biochemistry, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China
^b Department of Cardiology, Chongqing First-aid Medical Center, Chongqing 400014, China
^c Department of Ophthalmology, The Military General Hospital of Beijing PLA, Beijing 100700, China
^d Department of Dermatology, Feinberg School of Medicine, Northwestern University, Chicago 60611, USA

收稿日期:2010-05-05 出版日期:2010-06-10 发布日期:2010-06-10

Antagomir Dependent MicroRNA-205 Reduction Enhances Adhesion Ability of Human Corneal Epithelial Keratinocytes

Jun Li^1,2†, Hua Bai^3†, Yong Zhu¹, Xiao-yan Wang¹, Fang Wang¹, Jun-wu Zhang¹, Robert M. Lavker⁴, and Jia Yu^1*

^a Department of Biochemistry, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China
^b Department of Cardiology, Chongqing First-aid Medical Center, Chongqing 400014, China
^c Department of Ophthalmology, The Military General Hospital of Beijing PLA, Beijing 100700, China
^d Department of Dermatology, Feinberg School of Medicine, Northwestern University, Chicago 60611, USA

Received:2010-05-05 Online:2010-06-10 Published:2010-06-10
About author:Corresponding author Tel: 86-10-65296423, E-mail: j-yu@ibms.pumc.edu.cn

摘要/Abstract

摘要： 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. Conclusion Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.

关键词: corneal epithelium, cell signaling, microRNA

Abstract: 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. Conclusion Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.

Key words: corneal epithelium, cell signaling, microRNA

基金资助:

Supported by Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences Grant (National Institutes of Health Grants (Corresponding author Tel: 86-10-65296423

Jun Li, Hua Bai, Yong Zhu, Xiao-yan Wang, Fang Wang, Jun-wu Zhang, Robert M. Lavker and Jia Yu. Antagomir Dependent MicroRNA-205 Reduction Enhances Adhesion Ability of Human Corneal Epithelial Keratinocytes[J].Chinese Medical Sciences Journal, 2010, 25(2): 65-70.

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Antagomir Dependent MicroRNA-205 Reduction Enhances Adhesion Ability of Human Corneal Epithelial Keratinocytes

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