摘要
目的探讨缺氧条件下雷帕霉素促进人骨肉瘤MG63细胞凋亡是否与mTORC1/HIF-1α信号通路相关。方法流式细胞技术检测人骨肉瘤MG63细胞凋亡率;Western blot检测人骨肉瘤MG63细胞TSC1、p-S6、HIF-1α、Bax、Bcl-2等蛋白表达;免疫荧光检测人骨肉瘤MG63细胞HIF-1α蛋白表达。结果缺氧条件下,雷帕霉素促进人骨肉瘤MG63细胞凋亡,抑制人骨肉瘤MG63细胞mTORC1活性和HIF-1α蛋白表达;过表达HIF-1α能够抑制雷帕霉素对人骨肉瘤MG63细胞的促凋亡作用;mTORC1通过调控HIF-1α表达进而调节人骨肉瘤MG63细胞凋亡。结论缺氧条件下雷帕霉素可通过抑制mTORC1/HIF-1α信号通路促进人骨肉瘤MG63细胞凋亡。
Objective To explore whether rapamycin promote the apoptosis in human osteosarcoma MG63 cells under hypoxia is related to mTORC1/HIF-1αsignal pathway.Methods The apoptosis rate of MG63 cells was detected by flow cytometry,the protein expression of TSC1,p-S6,HIF-1α,Bax and Bcl-2 in MG63 cells were detected by Western blot,and the protein expression of HIF-1αin MG63 cells was detected by immunofluorescence.Results Under hypoxia condition,the increased apoptosis rate,the inhibition of mTORC1 activity and HIF-1αprotein expression in MG63 cells regulated by rapamycin were found.Overexpression of HIF-1αrestrained the proapoptotic effect of rapamycin in MG63 cells;mTORC1 regulated HIF-1αexpression and then regulated apoptosis of human osteosarcoma MG63 cells.Conclusions Rapamycin promotes MG63 apoptosis by inhibiting mTORC1/HIF-1αsingnal pathway uder hypoxia condition.
作者
王华磊
叶向阳
汤立新
WANG Hua-lei;YE Xiang-yang;TANG Li-xin(Dep of Orthopaedics,Nanyang Central Hospital,Nanyang,Henan 473000,China)
出处
《临床骨科杂志》
2020年第6期899-904,共6页
Journal of Clinical Orthopaedics
