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Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
Published on: January 7, 2013
Expression of protein kinase C isoforms in cultured human Tenon's capsule fibroblast cells
Ping Ma1, Qianying Gao2, Zhichong Wang2
1Department of Ophthalmology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, P.R. China.
Abstract:
Members of the protein kinase C (PKC) family are involved in physiological and pathophysiological processes, and exert an important role in signal transduction. The aim of the present study was to determine which of the 12 protein kinase C (PKC) isoforms (PKCα, PKCβⅠ, PKCβⅡ, PKCγ, PKCδ, PKCε, PKCη, PKCθ, PKCµ, PKCζ, PKCλ and PKCι) were expressed in vitro in cultured human Tenon's capsule fibroblasts (HTFs). HTFs from cell passages three to five were investigated for the presence of the 12 PKC isoforms at the cellular, mRNA and protein levels using laser scanning confocal microscopy (LSCM), reverse transcription‑polymerase chain reaction (RT‑PCR) and western blot analysis, respectively. These analyses yielded similar results for several of the PKC isoforms (PKCα, PKCδ, PKCε, PKCη, PKCµ, PKCζ, PKCλ and PKCι); however, PKCβⅠ, PKCβⅡ, PKCγ and PKCθ were not expressed in the cultured HTFs. LSCM revealed that eight PKC isoforms, PKCα, PKCδ, PKCε, PKCη, PKCµ, PKCζ, PKCλ and PKCι, appeared almost exclusively in the cytoplasm of the cells. Notably, PKCδ was expressed particularly well in the cytoskeleton. The present study revealed that all 12 PKC isoforms were expressed and that eight of the isoforms were present in cultured HTFs. These results will be of value when determining specific roles for the PKC isoforms in HTF proliferation, which may provide a novel therapeutic target for bleb scarring in glaucoma filtering surgery.
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