@article{APS4452,
author = {Yu-ping Zheng and Hui Liu and Hao Zeng and Lei Xiong and Zhao-hui Feng and Nai-xue Sun},
title = {Downregulation of lentivirus-mediated ILK RNAi on tractional force generation in human retinal Müller cells},
journal = {Acta Pharmacologica Sinica},
volume = {30},
number = {12},
year = {2016},
keywords = {},
abstract = {Aim: To investigate the effect of lentivirus-mediated integrin-linked kinase (ILK) RNA interference (RNAi) on human retinal Müller cells transdifferentiation into contractile myofibroblasts.
Methods: A lentiviral vector expressing ILK-specific shRNA was constructed and introduced into cultured retinal Müller cells. Silencing of the ILK gene was identified by real time RT-PCR and Western blot. The Müller cell phenotype change was confirmed by immunodetection of α-smooth muscle actin (α-SMA) stress fiber formation. The generation of tractional force was assessed using a tissue culture assay with cells incubated in three-dimensional collagen gels; cell migration was determined by the Boyden chamber method, using 10% FBS as a chemotactic factor.
Results: Significant decreases in ILK mRNA and protein expression were detected in Müller cells carrying lentiviral ILK-shRNA vector. Cells treated with anti-ILK siRNA showed less α-SMA stress fiber formation under hypoxic conditions or cell subcultivation. Lentiviral ILK-shRNA vector transfection also significantly reduced cell migration and cell-mediated gel contraction.
Conclusion: Lentivirus-mediated ILK RNAi decreased cell migration and contractile force generation by inhibiting α-SMA stress fiber formation in human retinal Müller cells. This tool might be useful to treat ocular fibroproliferative diseases associated with transdifferentiated Müller cells.},
issn = {1745-7254}, url = {http://www.chinaphar.com/article/view/4452}
}