蟾毒灵诱导骨肉瘤细胞株凋亡的蛋白质组学研究(4)
2. State Key Laboratory of Oncology in Southern， Sun Yat-sen University Cancer Center， Guangzhou 510060， China)
[Abstract] Objective： To study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.Method： MTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-2OS， U-2OS/MTX300， SaOS-2， IOR/OS9 treated with bufalin in different concentrations and times.The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then， the proteomic results were validated by western blot. Result： Bufalin inhibited the growth of human osteosarcoma cells U2OS，U2OS/MTX300 (methotrexate resistant cells)，SAOS2，IOR/OS9 in a dose- and time-dependent manner.The 72 h IC50 were (37.43±4.1)，(32.24±5.3) nmol·L-1 in U2OS，U2OS/MTX300 cells，respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins， the level of an anti-apoptotic protein， heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U2OS and U2OS/MTX300 cells. Conclusion： Bufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them， downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.
[Key words] bufalin； osteosarcoma； apoptosis； heat shock protein 27
doi：10.4268/cjcmm20141428
[责任编辑 陈玲], 百拇医药(谢显彪 温丽丽 尹军强 廖鸿益 邹昌业 王博 黄纲 沈靖南)
[Abstract] Objective： To study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.Method： MTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-2OS， U-2OS/MTX300， SaOS-2， IOR/OS9 treated with bufalin in different concentrations and times.The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then， the proteomic results were validated by western blot. Result： Bufalin inhibited the growth of human osteosarcoma cells U2OS，U2OS/MTX300 (methotrexate resistant cells)，SAOS2，IOR/OS9 in a dose- and time-dependent manner.The 72 h IC50 were (37.43±4.1)，(32.24±5.3) nmol·L-1 in U2OS，U2OS/MTX300 cells，respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins， the level of an anti-apoptotic protein， heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U2OS and U2OS/MTX300 cells. Conclusion： Bufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them， downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.
[Key words] bufalin； osteosarcoma； apoptosis； heat shock protein 27
doi：10.4268/cjcmm20141428
[责任编辑 陈玲], 百拇医药(谢显彪 温丽丽 尹军强 廖鸿益 邹昌业 王博 黄纲 沈靖南)