Unknown,Transcriptomics,Genomics,Proteomics

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THE GENE EXPRESSION PROFILE CHANGES OF HUMAN MEIBOMIAN GLAND EPITHELIAL CELLS UNDER DIFFERENTIATION

ABSTRACT: Analysis of gene expression profile changes of immortalized human meibomian gland epithelial cells under differentiation. Results provide important information of the response of human meibomian gland epithelial cells to different differentiation media, such as serum containing medium, serum-free medium, serum-free medium with 1.2mM Ca2+. Primary cultured human meibomian gland epithelial cells and hTERT immortalized human meibomian gland epithelial cells (3 cells/condition/experiment) were grown in keratinocyte serum-free medium (SFM, Invitrogen-Gibco, Grand Island, NY) on 75cm2 to reach confluence, then change to SFM, SFM with 1.2mM Calcium, or serum containing medium (10% fetal bovine serum in an equal volume of DMEM and Ham’s F12 with 10ng/ml EGF) for 14 days. Total RNA was extracted from the above cultures. Gene expression was analyzed using Illumina HumanHT-12 v3 Expression Beadchips (San Diego, CA).

ORGANISM(S): Homo sapiens

SUBMITTER: David Sullivan

PROVIDER: E-GEOD-23571 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:OBJECTIVE: The liver X receptor M-NM-1 (LXRM-NM-1) is a ligand-dependent nuclear receptor and the major regulator of reverse cholesterol transport in macrophages. This makes it an interesting target for mechanistic study and treatment of atherosclerosis. METHODS AND RESULTS: We optimized a promising stilbenoid structure (STX4) in order to reach nanomolar effective concentrations in LXRM-NM-1 reporter-gene assays. STX4 displayed the unique property to activate LXRM-NM-1 effectively but not its subtype LXRM-NM-2. The potential of STX4 to increase transcriptional activity as an LXRM-NM-1 ligand was tested with gene expression analyses in THP1-derived human macrophages and oxLDL-loaded human foam cells. Only in foam cells but not in macrophage cells STX4 treatment showed athero-protective effects with similar potency as the synthetic LXR ligand T0901317 (T09). Surprisingly, combinatorial treatment with STX4 and T09 resulted in an additive effect on reporter-gene activation and target gene expression. In physiological tests the cellular content of total and esterified cholesterol was significantly reduced by STX4 without the undesirable increase in triglyceride levels as observed for T09. CONCLUSIONS: STX4 is a new LXRM-NM-1-ligand to study transcriptional regulation of anti-atherogenic processes in cell or ex vivo models, and provides a promising lead structure for pharmaceutical development. 2 treatment groups (STX4 vs. DMSO in foam cells), 3 biological replicates per treatment (6 samples in total), each representing one well from cell culture plate 2 treatment groups (STX4 vs. DMSO in macrophages), 4 biological replicates per treatment (8 samples in total), each representing one well from cell culture plate 2 treatment groups (T0901317 vs. DMSO in foam cells), 3-4 biological replicates per treatment (7 samples in total), each representing one well from cell culture plate 2 treatment groups (T0901317 vs. DMSO in macrophages), 4 biological replicates per treatment (8 samples in total), each representing one well from cell culture plate

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THE GENE EXPRESSION PROFILE CHANGES OF HUMAN MEIBOMIAN GLAND EPITHELIAL CELLS UNDER DIFFERENTIATION

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