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Y. Khalifa, D. Xie, G. I. Liou, D. Bogorad, W. Bollag; Novel Lipid Signaling Pathway Modulates Corneal Epithelial Proliferation. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4308. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
SV40-immortalized human corneal epithelial cells were cultured in a 1:1 mixture of defined keratinocyte serum-free (dKSF) medium (Gibco) and minimum essential medium (Cellgro) until approximately 70% confluent. These near-confluent cells were then treated for 24 hours with dKSF medium containing increasing concentrations of PG liposomes, prepared via bath sonication of PG in dKSF. [3H]Thymidine incorporation into DNA was then determined. In separate experiments, near-confluent cells were treated for 24 hours with glycerol (0.1 and 0.5%) and/or calcium (1 mM versus a control concentration of 90 µM), and [3H]thymidine incorporation into DNA was then determined.
PG liposomes inhibited 3H]thymidine incorporation into DNA in a concentration-dependent manner, inducing a significant approximately 50% inhibition of DNA synthesis at 100 µg/mL. Glycerol (0.1 and 0.5%) and elevated extracellular calcium concentration (1 mM versus a control calcium concentration of 90 µM) more efficiently inhibit DNA synthesis than either agent alone.
PG liposomes inhibit the proliferation of SV40-immortalized human corneal epithelial cells similar to previous reports in mouse keratinocytes. We have also found that PLD2 and AQP3 localized to the same fractions upon sucrose density ultracentrifugation of SV40-immortalized human corneal epithelial cells, suggesting the likelihood that these two proteins may interact in corneal epithelial cells, as in epidermal keratinocytes. Our finding that glycerol together with an elevated extracellular calcium level and PG liposomes inhibit proliferation in a corneal epithelial cell model, as well as a report that AQP3 knockout mice exhibit delayed corneal wound healing, indicates that this novel signaling module might be critically involved in other corneal epithelial cell responses, such as corneal wound healing.
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