Moxifloxacin induces random migration in human corneal fibroblasts via the protein kinase C epsilon/zonula occludens-1 signaling pathway

Moxifloxacin (MOX) suppresses cell movement in human corneal fibroblasts (HCFs). Zonula occludens-1 (ZO-1) is localized to the leading edge of migrating HCFs. This study explored the role of ZO-1 in MOX-suppressed cell migration in HCFs. A single-cell trajectory analysis revealed that MOX negatively regulated the migratory properties of HCFs including migration distance, migration velocity, and directionality (P < 0.001, P < 0.001, and P = 0.018, respectively). MOX increased endogenous ZO-1 in HCFs in a concentration-dependent manner (P = 0.083, P = 0.005, and P = 0.001 at 10, 50, and 100 mu g/ml, respectively), but decreased the phosphorylation of endogenous ZO-1 at serines, threonines, and tyrosines. In contrast, MOX did not alter the expression of protein kinase C epsilon (PKCe), Rac-1, Cdc42, and MRCK8. However, MOX did also reduce the phosphorylation level of PKCe at serines and threonines (P < 0.001 at 100 mu g/ml). In addition, MOX increased the phosphorylation level of Rac-1 in a concentration-dependent manner (P < 0.001 at 100 mu g/ml). Compared with the mock cells, the directionality of cell movement increased significantly in ZO-1-expressing HCFs (P = 0.012) and decreased significantly in ZO-1-silenced HCFs (P = 0.002). The directionality did not change significantly in Rac-1-silenced HCFs. ZO-1-expressing HCFs moved faster than mock cells. PKCe, Cdc42, Rac-1, and phosphorylated Rac-1 were decreased in ZO-1-overexpressing HCFs, but increased in ZO-1-silenced HCFs. Finally, silencing ZO-1 blocked MOX hyperactivation of Rac-1. These suggest that MOX might trigger random migration in human corneal stromal cells through PKCe-modulated ZO-1 inactivation and Rac-1 hyperactivation.

Automated summary

The study revealed that MOX suppresses cell migration in HCFs by modulating ZO-1 and Rac-1 protein levels, indicating the underlying mechanisms in corneal stromal cells.