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- Title
Fibronectin and Focal Adhesion Kinase Small Interfering RNA Modulate Rat Retinal Müller Cells Adhesion and Migration.
- Authors
Xin-Ling Wang; Tao Yu; Jin-Song Zhang; Qi-Chang Yan; Ya-Hong Luo
- Abstract
Abstract Retinal Müller cells (RMCs) hypertrophy and proliferation play a crucial role in epiretinal membrane formation. This study was designed to analyze the effects of Fibronectin and specific FAK siRNA in cell adhesion and migration in rat Müller cells. RMCs were cultured and identified by GFAP, Vimentin, and GLAST mAb, respectively. The cells were planted on dishes coated with Fibronectin at 0, 1, 5, 10, 50, and 100 μg/ml. The attachment and migration assay was applied to characterize the RMCs–Fibronectin interactions. Cell lysis and Western blotting were utilized to detect β1-integrin, FAK, and GLAST protein expression. Then the cells were treated with FAK siRNA, non-targeting siRNA, and control medium. The cell cycle and apoptosis rate was determined by flow cytometry. The attachment, migration, and Western blotting assay were repeated. These data suggested that almost all the cells expressed GFAP, Vimentin, and GLAST, respectively, which ensured most of the harvested cells were RMCs. In attachment assay, the A570 values increased significantly with time (F = 1105.439, P F = 424.683, P F = 34.703, P 1-integrin, and GLAST elevated significantly as Fibronectin concentration increased (F = 54.755, P F = 119.962, P F = 39.287, P 0/G1 percentage and apoptosis rate compared with NT siRNA and control group (F = 11.526, P = 0.009; F = 64.772, P F = 10.500, P = 0.011). The A570 values were significantly suppressed in FAK siRNA groups compared with NT siRNA and control group (F = 154.241, P F = 10.906, P = 0.001). FAK and GLAST expression ratio decreased significantly after FAK siRNA treatment (F = 5.315, P = 0.047; F = 5.985, P = 0.042). Take together, FAK is involved in β1-integrin mediated adhesive signaling and play a critical role in regulating Müller cell adhesion, migration, and so far as to glutamate transportation functions.
- Subjects
FIBRONECTINS; FOCAL adhesion kinase; SMALL interfering RNA; LABORATORY rats; CELL adhesion; CELL migration; RETINA cytology; HYPERTROPHY; CELL proliferation
- Publication
Cellular & Molecular Neurobiology, 2009, Vol 29, Issue 4, p549
- ISSN
0272-4340
- Publication type
Article
- DOI
10.1007/s10571-009-9346-x