ABSTRACT
BACKGROUND: Emerging evidence indicates that the interaction between glucocorticoid receptor α (GRα) and nuclear factor κB (NF-κB) is a key pathogenetic cross talk in the autoimmune and inflammatory disorders. The objective of this study was to determine the GRα expression in patients with oral lichen planus (OLP) and investigate its correlation with NF-κB in OLP. METHODS: We compared the expression of GRα and NF-κB in oral biopsy specimens from patients with OLP(n = 32) against normal controls (n = 12) and investigated the correlation between the expression of GRα and NF-κB in OLP. RESULTS: Immunohistochemistry showed that GRα mainly expressed in the cytoplasm of keratinocytes of basal and spinosum layer of OLP. Both real-time quantitative PCR and Western blots revealed that the mRNA and protein expression levels of GRα were decreased compared with normal controls (both P < 0.001). Conversely, those levels of nuclear factor-kappa B (NF-κB) were increased compared with normal controls (both P < 0.001). Importantly, a significant inverse correlation between the GRα and NF-κB was found (P < 0.05). CONCLUSIONS: Our findings demonstrated that low expression of GRα in OLP correlates with activation of NF-κB, which indicates that the cross talk between GRα and NF-κB in OLP may become a new therapeutic target and represent a new approach to explore the pathogenesis of OLP.
Subject(s)
Lichen Planus, Oral/metabolism , NF-kappa B/analysis , Receptors, Glucocorticoid/analysis , Adult , Blotting, Western , Case-Control Studies , Cell Nucleus/pathology , Cytoplasm/pathology , Female , Humans , Immunohistochemistry , Keratinocytes/pathology , Lichen Planus, Oral/pathology , Male , Middle Aged , Mouth Mucosa/pathology , NF-kappa B p50 Subunit/analysis , Real-Time Polymerase Chain Reaction , Receptor Cross-Talk/physiology , Transcription Factor RelA/analysis , Young AdultABSTRACT
OBJECTIVES: The aim of this study was to establish a stable in vitro culture system for keratinocytes obtained from oral lichen planus (OLP) lesions and evaluate cultured keratinocyte characteristics including cell morphology, ultrastructure, and expression of biomarkers. MATERIALS AND METHODS: OLP mucosa (histopathologically confirmed) was collected and cells isolated using the cold enzyme digestion method. Primary culture and serial passage were performed on serum-free keratinocyte medium. Morphological changes of cells were evaluated via inverted phase contrast microscopy, and cellular ultrastructure was observed by electron microscopy. Indirect immunofluorescence was used to detect expression of keratin and nuclear factor-kappaB (NF-κB). RESULTS: OLP type I keratinocytes was successfully cultured in vitro in serum-free medium. Cellular morphology was typically polygonal during the growth phase. Cells could be passaged continuously for five to six generations without losing viability. Transmission electron microscopy showed large nuclei and multiple vacuoles in the cultured cells consistent with histopathological features of OLP keratinocytes. Indirect immunofluorescence staining was positive for keratin and NF-κB. CONCLUSIONS: This study established that human OLP kera-tinocytes can be successfully cultured cells with histopathologic features and biomarker expression consistent with OLP type I keratinocytes. CLINICAL RELEVANCE: This culture system lays a foundation for the establishment of human OLP cell model in vitro.
