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Androgen regulation of gene expression in human meibomian gland and conjunctival epithelial cells.

ABSTRACT: Androgens exert a significant influence on the structure, function and/or pathophysiology of the meibomian gland and conjunctiva. We sought to determine whether this hormone action involves the regulation of epithelial cell gene expression in these tissues.Immortalized human meibomian gland and conjunctival epithelial cells were treated with placebo or dihydrotestosterone (DHT) and processed for molecular biologic procedures. Gene expression was evaluated with BeadChips and data were analyzed with bioinformatic and statistical software.Androgen treatment significantly influenced the expression of approximately 3,000 genes in immortalized human meibomian gland and conjunctival epithelial cells. The nature of DHT action on gene activity was predominantly cell-specific. Similarly, DHT exerted a significant, but primarily cell-specific, influence on many gene ontologies and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. These included groups of genes related, for example, to lipid dynamics, innate immunity, cell cycle, Janus kinase (JAK)-signal transducer and activator of transcription (stat) cascades, oxidative phosphorylation, the proteasome, and mammalian target of rapamycin (mTOR), Wnt, and peroxisome proliferator-activated receptor (PPAR) signaling.Our findings support our hypothesis that androgens regulate gene expression in human meibomian gland and conjunctival epithelial cells. Our ongoing studies are designed to determine whether many of these genes are translated and play a role in the health and well being of the eye.

SUBMITTER: Khandelwal P

PROVIDER: S-EPMC3351406 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Androgen regulation of gene expression in human meibomian gland and conjunctival epithelial cells.

Khandelwal Payal P Liu Shaohui S Sullivan David A DA

Molecular vision 20120427

<h4>Purpose</h4>Androgens exert a significant influence on the structure, function and/or pathophysiology of the meibomian gland and conjunctiva. We sought to determine whether this hormone action involves the regulation of epithelial cell gene expression in these tissues.<h4>Methods</h4>Immortalized human meibomian gland and conjunctival epithelial cells were treated with placebo or dihydrotestosterone (DHT) and processed for molecular biologic procedures. Gene expression was evaluated with Bea ...[more]

PMID: 22605918

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Project description:Human conjunctival cell lines are useful tools for modeling ocular surface disease and evaluation of ocular drugs and cosmetics. However, gene expression in these cells may not be comparable to primary cultured cells, raising doubts that they could be used as a substitute. We aimed to ascertain the similarities of global gene expression between commonly used cell lines and primary cells using a microarray approach. The Affymetrix U133A chip (>22,000 genes) was used to investigate conjunctival tissue (CT), primary conjunctival epithelial cells (PCEC), two conjunctival epithelial cell lines (IOBA-NHC and ChWK), and HCEC-T, a human corneal epithelial cell line (control). Using principal component analysis, the PCEC profile was clustered more closely to conjunctival tissue than either of the two cell lines. Certain extracellular matrix genes were differentially upregulated in CT compared to PCEC, suggesting presence of fibroblasts in addition to epithelial cells in CT. Overall, 67.3% (95% CI: 66.7-67.9) of transcripts in IOBA-NHC were within 1.5-fold of the corresponding transcripts in PCEC, but only 62.2% (95% CI: 61.5-62.9) in the case of ChWK. In HCEC-T, the proportion was only 58.8% (95% CI 58.1-59.4), suggesting less resemblance to PCEC than the conjunctival epithelial cell lines. The IOBA-NHC profile was more similar to PCEC than ChWK, for all genes and genes concerned with membrane association, communication, development, and regulation of metabolism, especially protein and nucleic acid metabolism. The correlation of normalized gene expression levels was high between either the IOBA-NHC or ChWK and PCEC for genes concerned with cell defense, viral life cycle, antigen presentation, antioxidation, or ubiquitin ligation. In order to evaluate the functional significance of the altered gene expression in IOBA-NHC cells, we evaluated a few proteins important for epithelial differentiation or defense, corresponding to the transcripts for S100A9, TGM2, and TLR4. Protein levels of S100A9 and TGM2 were indeed raised, and TLR4 decreased, in IOBA-NHC compared to PCEC. Gene expression in conjunctival cell lines differs from primary cells, but the profile varies according to functional gene categories. Depending on the methodology of proposed studies, if there is limited availability of PCEC, NHC-IOBA may be more suitable than ChWK, but even then, epithelial differentiation and innate immunity functions in NHC-IOBA may differ from primary cells.

Dataset Information

Androgen regulation of gene expression in human meibomian gland and conjunctival epithelial cells.

Publications

Androgen regulation of gene expression in human meibomian gland and conjunctival epithelial cells.

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