title: Microarray analysis of 1,25(OH)₂D₃ regulated gene expression in human primary osteoblasts.
abstract: Though extensive studies have been conducted, questions regarding the molecular effectors and pathways underlying the regulatory role of 1,25(OH)(2)D(3) in human osteoblasts other than cell differentiation and matrix protein production remain unanswered. This study aims to identify genes and pathways that are modulated by 1,25(OH)(2)D(3) treatment in human osteoblasts. Primary osteoblast cultures obtained from human bone tissue samples were treated with 1,25(OH)(2)D(3) (10(-7)  M) for 24 h and their transcritptomes were profiled by microarray analysis using the Affymetrix GeneChip. Statistical analysis was conducted to identify genes whose expression is significantly modulated following 1,25(OH)(2)D(3) treatment. One hundred and fifty-eight genes were found to be differentially expressed. Of these, 136 were upregulated, indicating clear transcriptional activation by 1,25(OH)(2)D(3). Biostatistical evaluation of microarray data by Ingenuity Pathways Analysis (IPA) revealed a relevant modulation of genes involved in vitamin D metabolism (CYP24), immune functions (CD14), neurotransmitter transporters (SLC1A1, SLC22A3), and coagulation [thrombomodulin (THBD), tissue plasminogen activator (PLAT), endothelial protein C receptor (PROCR), thrombin receptor (F2R)]. We identified a restricted number of highly regulated genes and confirmed their differential expression by real-time quantitative PCR (RT qPCR). The present genome-wide microarray analysis on 1,25(OH)(2)D(3) -treated human osteoblasts reveals an interplay of critical regulatory and metabolic pathways and supports the hypothesis that 1,25(OH)(2)D(3) can modulate the coagulation process through osteoblasts, activates osteoclastogenesis through inflammation signaling, modulates the effects of monoamines by affecting their reuptake.
authors.affiliation: Axxam Spa, San Raffaele Biomedical Science Park, 20132 Milan, Italy. paola.tarroni.pt@axxam.com
authors.forename: Paola
authors.initials: P
authors.lastname: Tarroni
authors.forename: Isabella
authors.initials: I
authors.lastname: Villa
authors.forename: Emanuela
authors.initials: E
authors.lastname: Mrak
authors.forename: Francesca
authors.initials: F
authors.lastname: Zolezzi
authors.forename: Michela
authors.initials: M
authors.lastname: Mattioli
authors.forename: Claudio
authors.initials: C
authors.lastname: Gattuso
authors.forename: Alessandro
authors.initials: A
authors.lastname: Rubinacci
mesh_terms: D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D002117:Calcitriol / Q000494:pharmacology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D010006:Osteoblasts / Q000378:metabolism*; D061251:Primary Cell Culture; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D014815:Vitamins / Q000494:pharmacology*
chemical_list: D000943:Antigens, Differentiation; D014815:Vitamins; D002117:Calcitriol