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Purpose To investigate the autophagy characteristics of giant cell tumor of bone and its effect on cell proliferation. Methods Giant cell tumors of bone (GCT-0404 cells) were cultured in vitro and treated with serum-starvation, rapamycin and inhibitor chloroquine. Western blot analysis was performed to detect the autophagic markers LC3 and Beclin 1 expression. Immunofluorescence method was used to show intracellular autophagy formation. Inverted microscope was used to observe the cell morphology. CCK-8 assays were used to detect the cell viability. Results Green fluorescent spots that represented the transformation level of LC3-I to LC3-Ⅱ increased significantly (P＜0.05) by serum-starvation, rapamycin and inhibitor chloroquine respectively, but the expression level of Beclin 1 was not raised. In addition, the cell morphology changed significantly, and cell proliferation was inhibited after serum-starvation, and treatment with rapamycin and inhibitor chloroquine (P＜0.05), Conclusion The changes of autophagy caused by serum starvation, rapamycin and inhibitor chloroquine in GCT-0404 cells may be involved in the down-regulation of cell proliferation.
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Objective To observe the effects of genistein and 17?-estradiol on microstructure of cancellous bone in ovariectomized (OVX) rats.Methods Ninty 7-month-old SD rats were randomly divided into baseline group,ovariectomized (OVX),sham-operated (SHAM),17?-estradiol treated (10?g?kg~(-1).day~(-1),EST) and genistein treated (5 mg?kg~(-1)?day~(-1),GEN) groups,and were killed at the beginning of the experiment,the 3rd and 15th week after operation.MicroCT scanning was performed on the left tibia in vitro.The regions involving 0.5 mm slice thickness and 1.6 mm distal to the tibial growth plate were selected as the regions of interest.Results At the 3rd week after operation,the tissue bone mineral density (tBMD) and trabecular thickness (sTh.Th) in group GEN were significantly higher than those in OVX and EST groups (all P