龙葵碱对骨肉瘤细胞放射敏感性的影响

Effect of solanine on radiosensitivity of osteosarcoma cells

目的研究龙葵碱对骨肉瘤细胞放射敏感性的影响。方法用龙葵碱处理骨肉瘤细胞,同时给予6 MV X射线照射,噻唑蓝(MTT)方法测定细胞增殖变化,流式细胞术测定细胞凋亡,Western blot检测细胞中Ki-67、 PCNA、 Bax、 Cleaved Caspase-3蛋白水平,JC-1法检测线粒体膜电位,DCFH-DA法检测细胞中ROS水平,平板克隆实验测定放射敏感性。结果与未经龙葵碱和照射后的细胞比较,龙葵碱或照射处理后骨肉瘤细胞增殖能力和Ki-67、 PCNA蛋白水平均降低(F=55.165、 50.667、 23.389,P<0.05),细胞凋亡率和Bax、 Cleaved Caspase-3蛋白水平均升高(F=54.588、 42.924、 51.541,P<0.05),线粒体膜电位下降(F=40.762,P<0.05),细胞中ROS水平升高(F=79.055,P<0.05)。与龙葵碱或单纯照射的细胞比较,龙葵碱联合照射处理后的骨肉瘤细胞增殖能力和Ki-67、 PCNA蛋白水平降低(F=55.165、 50.667、 23.389,P<0.05),细胞凋亡率和Bax、 Cleaved Caspase-3水平升高(F=54.588、 42.924、 51.541,P<0.05),线粒体膜电位降低(F=40.762,P<0.05),ROS水平升高(F=79.055,P<0.05)。龙葵碱对骨肉瘤细胞的放射增敏比为1.786。结论龙葵碱抑制骨肉瘤细胞增殖并诱导凋亡,提高骨肉瘤细胞放射敏感性。

Objective To investigate the effect of solanine on radiosensitivity of osteosarcoma cells. Methods Osteosarcoma cells were treated with solanine, at the same time, radiation treatment was given, cell proliferation was measured by MTT assay, cell apoptosis was measured by flow cytometry, the protein levels of Ki-67, PCNA, Bax and Cleaved Caspase-3 were detected by Western blot and mitochondrial membrane potential was detected by JC-1 method, the ROS level in cells was detected by DCFH-DA method. Radiosensitivity was measured by plate cloning. Results Compared with cells that were not treated with solanine and radiation, the proliferation of osteosarcoma cells and the Ki-67 and PCNA protein levels were decreased after solanine or radiotherapy (F=55.165, 50.667, 23.389, P<0.05), the apoptotic rate and the level of Bax and Cleaved Caspase-3 protein increased (F=54.588, 42.924, 51.541, P<0.05), the mitochondrial membrane potential decreased (F=40.762, P<0.05), and the ROS level in the cells increased(F=79.055, P<0.05). Compared with cells treated with solanine or radiotherapy, the proliferation of osteosarcoma cells and the levels of Ki-67 and PCNA proteins were decreased after solanine combined with radiation treatment (F=55.165, 50.667, 23.389, P<0.05), the apoptotic rate and the levels of Bax and Cleaved Caspase-3 were increased (F=54.588, 42.924, 51.541, P<0.05), the mitochondrial membrane potential was decreased (F=40.762, P<0.05), and the ROS level was increased (F=79.055, P<0.05). The radiosensitization ratio of solanine to osteosarcoma cells was 1.786. Conclusions Solanine inhibits proliferation and induces apoptosis of osteosarcoma cells, and improves radiosensitivity of osteosarcoma cells.