Epigenetic silencing of the CIITA gene and posttranscriptional regulation of class II MHC genes in ocular melanoma cells

Michael Radosevich, Zhimin Song, Joan C. Gorga, Bruce Ksander, Santa Jeremy Ono

Research output: Contribution to journalArticlepeer-review

Abstract

PURPOSE. Primary uveal melanocytes and many ocular melanoma cells are resistant to interferon (IFN)-γ-mediated induction of major histocompatibility complex (MHC) class II molecule expression. This suppression of class II MHC induction is considered to be one of the ways in which the eye is able to inhibit inflammatory responses. However, the mechanism(s) of this suppression is unknown. In this study, we have probed the molecular basis of this phenotype and report two distinct mechanisms underlying this phenotype. METHODS. Primary ocular melanocytes and ocular melanoma cell lines (retaining this IFN-γ-resistant class II MHC phenotype) were examined for the expression of class II MHC molecules on the cell surface by flow cytometry. Class II MHC gene expression was further examined using Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR) analyses. RESULTS. The IFN-γ signal-transduction pathway was found to be intact by electrophoretic mobility shift assay (EMSA) and transfection of reporter constructs. The lack of class II MHC gene expression appears to result from at least two mechanisms: (1) a specific inhibition of CIITA (class II transactivator) gene expression (reminiscent of trophoblasts), and (2) posttranscriptional regulation of class II MHC genes. CONCLUSIONS. The inability of primary uveal melanocytes and ocular melanoma cells to express class II MHC molecules after treatment with IFN-γ has been found to map to two distinct points in the class II MHC biosynthetic pathway. The predominant mechanism appears to involve the silencing of the endogenous gene encoding the class II transactivator (CIITA). Here, the blockade does not involve signal transduction from the IFN-γ receptor, but rather involves a specific silencing of the CIITA gene. A second mechanism involves the posttranscriptional regulation of class II MHC genes.

Original languageEnglish (US)
Pages (from-to)3185-3195
Number of pages11
JournalInvestigative Ophthalmology and Visual Science
Volume45
Issue number9
DOIs
StatePublished - Sep 2004

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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