Attenuation of human lens epithelial cell spreading, migration and contraction via downregulation of the PI3K/Akt pathway

Abstract

Posterior capsule opacification (PCO) represents a major challenge in the postoperative management of cataract patients. Spreading, migration and contraction of residual human lens epithelial cells play a pivotal role in the pathogenesis of PCO. Therefore, we analyzed the effect of the alkylphosphocholine (APC) erufosine on these cellular features as well as on PI3K/Akt, a crucial pathway in PCO pathogenesis. Human lens epithelial cells were cultured under standard cell culture conditions. Cell spreading was analyzed on fibronectin-coated wells and chemokinetic migration was assessed by time-lapse microscopy. For evaluation of cell-mediated collagen matrix contraction, the cells were seeded into collagen gels and incubated with an APC in different non-toxic concentrations before the surface area was measured on day 6. The activity of PI3K/Akt was assessed by an ELISA kit after incubation of the cells with different APC concentrations. Human lens epithelial cell spreading and migration were attenuated by APCs as follows: 7% spreading, 48% migration (0.1μM APC), and 32% spreading, 68% migration (1.0μM APC). APC concentrations of 0.1μM reduced collagen gel diameter by 5%, and 1.0μM by less than 1%, compared to untreated, cell-populated gels that resulted in a cell diameter contraction of 36%. PI3K was downregulated in a concentration-dependent manner. The crucial cellular features of PCO pathogenesis are attenuated by the APC erufosine via downregulation of the PI3K pathway. Thus, erufosine might become a valuable tool for pharmacologic PCO prophylaxis in the future.