Effect Of Myocilin On Fibronectin Expression And Cell Adhesion And Migration In Human Trabecular Meshwork Cells

Purpose To investigate the effect of myocilin on fibronectin expression and cellattachment and migration on the basis of culturing the trabecular meshwork cells fromnormal eyes，to study the relationship between myocilin and POAG.Methods Human trabecular meshwork cells were primarily cultured and subcultured，and then identified by electronic microscope and immunohistochemistry. Thetrabecular meshwork cells were treated with0ug/ml(control group),0.5ug/ml,1ug/ml,1.5ug/ml,2ug/ml,2.5ug/ml recombinant myocilin which were diluted byDMEM/F12without serum for24hours. Western blot and Elisa were used to detectthe fibronectin expression in cultured trabecular meshwork cells. CCK-8assay andtranswell chamberin were used to detect adhesion and migration ability of thetrabecular meshwork cells.Rusults1. The trabecular meshwork cells were successfully cultured and identified invitro.2. Within a certain range,as the myocilin concentration increased,the fibronectinlevel were reduced.3.The fibronectin concentration of trabecular cells which weretreated with0ug/ml,0.5ug/ml,1ug/ml,1.5ug/ml,2ug/ml,2.5ug/ml recombinantmyocilin were0.5817±0.0959ug/ml,0.5572±0.044ug/ml,0.5868±0.0735ug/ml,0.5015±0.0472ug/ml,0.4263±0.0426ug/ml,0.2838±0.0972ug/ml by Elisa assay. The1.5ug/ml、2ug/ml、2.5ug/ml myocilin experimental group compared with the control groupand within these three experimental groups the difference were statisticallysignificant(P<0.05).4.Our results showed that there were more migrated cells incontrol group than in experimental group, and the difference between them wasstatistically significant.5. The OD value of trabecular meshwork cells which weretreated with0ug/ml,0.5ug/ml,1ug/ml,1.5ug/ml,2ug/ml,2.5ug/ml myocilin were0.1614±0.0061，0.1529±0.0139，0.1273±0.0172，0.1271±0.0230，0.1165±0.0084，0.1103±0.0045by CCK-8assay. As the myocilin concentration increased,the OD value werereduced. And the difference between experimental groups and control group werestatistically significant(P <0.05). Conclusion1.Myocilin could reduce the fibronectin expression in trabecularmeshwork cells, and showed a dose dependent manner.2.The migration and adhesionability of trabecular meshwork cells were inhibited by myocilin.