title: Differentially expressed genes and signalling pathways are involved in mouse osteoblast-like MC3T3-E1 cells exposed to 17-β estradiol.
abstract: Oestrogen is essential for maintaining bone mass, and it has been demonstrated to induce osteoblast proliferation and bone formation. In this study, complementary DNA (cDNA) microarrays were used to identify and study the expression of novel genes that may be involved in MC3T3-E1 cells' response to 17-β estradiol. MC3T3-E1 cells were inoculated in minimum essential media alpha (α-MEM) cell culture supplemented with 17-β estradiol at different concentrations and for different time periods. MC3T3-E1 cells treated with 10⁻⁸ mol⋅L⁻¹ 17-β estradiol for 5 days exhibited the highest proliferation and alkaline phosphatase (ALP) activity; thus, this group was chosen for microarray analysis. The harvested RNA was used for microarray hybridisation and subsequent real-time reverse transcription polymerase chain reaction (RT-PCR) to validate the expression levels for selected genes. The microarray results were analysed using both functional and pathway analysis. In this study, microarray analysis detected 5403 differentially expressed genes, of which 1996 genes were upregulated and 3407 genes were downregulated, 1553 different functional classifications were identified by gene ontology (GO) analysis and 53 different pathways were involved based on pathway analysis. Among the differentially expressed genes, a portion not previously reported to be associated with the osteoblast response to oestrogen was identified. These findings clearly demonstrate that the expression of genes related to osteoblast proliferation, cell differentiation, collagens and transforming growth factor beta (TGF-β)-related cytokines increases, while the expression of genes related to apoptosis and osteoclast differentiation decreases, following the exposure of MC3T3-E1 cells to α-MEM supplemented with 17-β estradiol. Microarray analysis with functional gene classification is critical for a complete understanding of complementary intracellular processes. This microarray analysis provides large-scale gene expression data that require further confirmatory studies.
authors.affiliation: Department of Prosthodontics, College of Stomatology, Shandong University, Jinan, China.
authors.forename: Zhen-Zhen
authors.initials: ZZ
authors.lastname: Shang
authors.affiliation: Department of Stomatology, The Affiliated Hospital of Medical College Qingdao University, Qingdao, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: 1] Department of Prosthodontics, College of Stomatology, Shandong University, Jinan, China [2] Shandong Provincial Key Laboratory of Oral Biomedicine, Shandong University, Jinan, China.
authors.forename: Hui-Qiang
authors.initials: HQ
authors.lastname: Sun
authors.affiliation: Department of Prosthodontics, College of Stomatology, Shandong University, Jinan, China.
authors.forename: Guo-Ning
authors.initials: GN
authors.lastname: Xiao
authors.affiliation: Department of Prosthodontics, College of Stomatology, Shandong University, Jinan, China.
authors.forename: Cun-Wei
authors.initials: CW
authors.lastname: Wang
authors.affiliation: Department of Prosthodontics, College of Stomatology, Shandong University, Jinan, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Gong
mesh_terms: D016475:3T3 Cells; D000469:Alkaline Phosphatase / Q000187:drug effects; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D003094:Collagen / Q000187:drug effects / Q000235:genetics; D004396:Coloring Agents; D016207:Cytokines / Q000187:drug effects / Q000235:genetics; D004958:Estradiol / Q000008:administration & dosage / Q000494:pharmacology*; D004967:Estrogens / Q000008:administration & dosage / Q000494:pharmacology*; D020869:Gene Expression Profiling; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D010006:Osteoblasts / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics*; D013778:Tetrazolium Salts; D013844:Thiazoles; D016212:Transforming Growth Factor beta / Q000187:drug effects / Q000235:genetics
chemical_list: D004396:Coloring Agents; D016207:Cytokines; D004967:Estrogens; D013778:Tetrazolium Salts; D013844:Thiazoles; D016212:Transforming Growth Factor beta; D004958:Estradiol; D003094:Collagen; D000469:Alkaline Phosphatase; C022616:thiazolyl blue