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Expression of Phospholipases A₂ and C in Human Corneal Epithelial Cells

¹From Unité de Recherche en Ophtalmologie and ⁵Centre de Recherche en Oncologie et Endocrinologie Moléculaire, Centre de Recherche du Centre Hospitalier Universitaire de Québec (CHUQ), Pavillon du Centre Hospitalier de l’Université Laval (CHUL), Faculté de Médecine; and ³Laboratoire d’Organogenèse Expérimentale, Centre Hospitalier Affilié Universitaire, Pavillon Saint-Sacrement, Université Laval, Sainte-Foy, Québec, Canada; and the ⁴Departments of Chemistry and Biochemistry, University of Washington, Seattle, Washington.

PURPOSE. To achieve a better understanding of the involvement of phospholipases in the inflammation and wound-healing processes in human corneal epithelial cells (HCECs), expression of phospholipase A₂s (PLA₂s) and phospholipase Cs (PLCs) was examined in the human corneal epithelium.

METHODS. Specific primers were designed for RT-PCR amplification of the known secreted (s)PLA₂, cytosolic (c)PLA₂, and PLC mRNAs. Corresponding PCR products were cloned and the DNA sequenced. Immunofluorescence of flatmounted corneal sections and Western blot analyses were used to detect the PLA₂s and PLCs expressed by HCECs.

RESULTS. The mRNAs for the following phospholipases were detected by RT-PCR in the HCECs: sPLA₂GIII, -GX, and -GXIIA; cPLA₂ and -; PLCß1, -ß2, -ß3, -ß4, -1, -2, -1, -3, -4, and -. Immunofluorescence analyses conducted on corneal epithelium cryosections and Western blot on freshly isolated HCECs demonstrated the presence of sPLA₂GIII, -GX, and -GXIIA; cPLA₂ and -; and PLCß2, -ß3, -1, -2, and -3.

CONCLUSIONS. Many phospholipase isoforms are expressed by HCECs and may play a major role in signal transduction (PLCs) as well as in the release of precursors of potent mediators of inflammation, such as leukotrienes and prostaglandins (PLA₂s). Moreover, the sPLA₂s expressed by the corneal epithelium could be involved in the normal antibacterial activity in the tears and in wound healing.

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