Downregulation of lentivirus-mediated ILK RNAi on tractional force generation in human retinal Müller cells
| Autor: | Hao Zeng, Nai-xue Sun, Lei Xiong, Hui Liu, Yu-Ping Zheng, Zhao-hui Feng |
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| Rok vydání: | 2009 |
| Předmět: |
Stress fiber
Cellular differentiation Genetic Vectors Cell Down-Regulation Protein Serine-Threonine Kinases Biology Retina Small hairpin RNA Cell Movement Stress Fibers medicine Humans Pharmacology (medical) Integrin-linked kinase Cells Cultured Pharmacology Lentivirus Transdifferentiation Cell Differentiation Muscle Smooth Cell migration General Medicine Transfection Fibroblasts Molecular biology Actins medicine.anatomical_structure embryonic structures biology.protein Original Article RNA Interference sense organs Neuroglia |
| Zdroj: | Acta Pharmacologica Sinica. 30:1625-1633 |
| ISSN: | 1745-7254 1671-4083 |
| DOI: | 10.1038/aps.2009.154 |
| Popis: | To investigate the effect of lentivirus-mediated integrin-linked kinase (ILK) RNA interference (RNAi) on human retinal Müller cells transdifferentiation into contractile myofibroblasts.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 alpha-smooth muscle actin (alpha-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.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 alpha-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.Lentivirus-mediated ILK RNAi decreased cell migration and contractile force generation by inhibiting alpha-SMA stress fiber formation in human retinal Müller cells. This tool might be useful to treat ocular fibroproliferative diseases associated with transdifferentiated Müller cells. |
| Databáze: | OpenAIRE |
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