Differentially expressed genes and signalling pathways are involved in mouse osteoblast-like MC3T3-E1 cells exposed to 17-β estradiol
Authors: Zhen-Zhen Shang, Xin Li, Hui-Qiang Sun, Guo-Ning Xiao, Cun-Wei Wang, Qi Gong
Journal: International Journal of Oral Science (2014)

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 5 403 differentially expressed genes, of which 1 996 genes were upregulated and 3 407 genes were downregulated, 1 553 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. Bone-forming cells drastically alter their gene expression patterns in response to the hormone estrogen, according to researchers in China. A team led by Hui-Qiang Sun from Shandong University used microarray analyses to identify all the genes in a mouse bone-forming cell line (MC3T3-E1), where expression increased or decreased following treatment with a form of estrogen known as 17-β estradiol. Approximately 5 400 genes showed changed expression patterns. The researchers classified these genes into more than 1 500 different functional groups involved in approximately 50 different molecular pathways. In general, genes associated with cell proliferation and differentiation were switched on by estrogen, whereas genes involved in programmed cell death and bone resorption were switched off by the hormone treatment. The findings could guide therapies that aim to promote bone regeneration.