Expression of Protein Kinase C in Human Tenon's Capsule Fibroblasts

Abstract

Purpose: : Protein kinase C (PKC) is involved in both physiological and pathophysiological processes and plays an important role in signal transduction. The aim of the present study was to determine twelve PKC isoenzymes(PKC_α, PKC_βI, PKC_βII, PKC_γ, PKC_Δ,PKC_ε, PKC, PKC_θ, PKC_µ, PKC_ζ,PKC and PKC) expressesd in cultured human Tenon’s capsule fibroblasts (HTFs).

Methods: : HTFs were cultured in DMEM/F12 medium supplemented with 10% fetal bovine serum, identified with the growing characteristics, morphological features. HTFs from passage two or three were examined for 12 isoforms at the cellular, mRNA and protein levels by laser scanning confocal microscope (LSCM), reverse transcription(RT)-PCR and Western blot analysis, respectively.

Results: : The HTFs showed fusiform shape with abundant cytoplasm, great nucleus, with vimentin positive while cytokeratin negative. LSCM, RT-PCR and Western blot analysis showed similar results for specific PKC isoforms namely PKC_α, PKC_Δ,PKC_ε, PKC, PKC_µ, PKC_ζ,PKC and PKC , but not PKC_βI, PKC_βII, PKC_γ and PKC_θ were constantly expressed in HTFs with the exception of PKC at mRNA level. Confocal microscopy showed that eight PKC isoforms PKC_α, PKC_Δ,PKC_ε, PKC, PKC_µ, PKC_ζ,PKC and PKC appeared almost exclusively in the cytoplasm of the cells.

Keywords: proliferation • signal transduction