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...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-b estradiol. MC3T3-E1 cells were inoculated in minimum essential media alpha（a-MEM）cell culture supplemented with 17-b estradiol at different concentrations and for different time periods. MC3T3-E1 cells treated with1028mol？L2117-b 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-b）-related cytokines increases, while the expression of genes related to apoptosis and osteoclast differentiation decreases, following the exposure of MC3T3-E1 cells to a-MEM supplemented with 17-b 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.展开更多
Objective:To investigate the role of SMB on the growth , differentiation and metabolism of osteoblastlike cells in vitro, we studied the effects of SMB on DNA synthesis and alkaline phosphatase(ALPase) activity of the...Objective:To investigate the role of SMB on the growth , differentiation and metabolism of osteoblastlike cells in vitro, we studied the effects of SMB on DNA synthesis and alkaline phosphatase(ALPase) activity of the cloned osteoblast like cells-MC3T3-E1. Methods: The cells were cultured in α-MEM with 0. 3% of fetal bovine serum and treated with SMB at the concentration of 0. 1 - 10. 0 g/L. Results : There was no significant difference in DNA synthesis between the groups in different concentration of SMB and the group of control. But SMB increased ALPase activity in a concentration-dependent fashion in later stage of cells , and up to maximum at the level of 5. 0 g/L , in which concentration , ALPase activity was about 135 % greater than that of control. However ,ALPase activity was inhibited in early stage of cells by the addition of SMB. Conclusion : SMB has the stimulating effect on the activity of osteoblast-like cells in vitro, and may play an important role in accelerating the remodeling of bone in vivo as well.展开更多
基金supported by grants from the Natural Science Fund (ZR2010HM035) of Shandong Provincethe Shandong Provincial Health Development Project Fund (2011WSB19002) in China
文摘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-b estradiol. MC3T3-E1 cells were inoculated in minimum essential media alpha（a-MEM）cell culture supplemented with 17-b estradiol at different concentrations and for different time periods. MC3T3-E1 cells treated with1028mol？L2117-b 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-b）-related cytokines increases, while the expression of genes related to apoptosis and osteoclast differentiation decreases, following the exposure of MC3T3-E1 cells to a-MEM supplemented with 17-b 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.
文摘Objective:To investigate the role of SMB on the growth , differentiation and metabolism of osteoblastlike cells in vitro, we studied the effects of SMB on DNA synthesis and alkaline phosphatase(ALPase) activity of the cloned osteoblast like cells-MC3T3-E1. Methods: The cells were cultured in α-MEM with 0. 3% of fetal bovine serum and treated with SMB at the concentration of 0. 1 - 10. 0 g/L. Results : There was no significant difference in DNA synthesis between the groups in different concentration of SMB and the group of control. But SMB increased ALPase activity in a concentration-dependent fashion in later stage of cells , and up to maximum at the level of 5. 0 g/L , in which concentration , ALPase activity was about 135 % greater than that of control. However ,ALPase activity was inhibited in early stage of cells by the addition of SMB. Conclusion : SMB has the stimulating effect on the activity of osteoblast-like cells in vitro, and may play an important role in accelerating the remodeling of bone in vivo as well.
