taurolidine对小鼠黑色素瘤细胞辐射敏感性的作用及机制

目的：研究taurolidine通过增强Bax和Bad蛋白表达，降低Bcl-2蛋白表达，提高小鼠黑色素瘤B16-F10细胞的辐射敏感性的作用及机制。方法：流式细胞仪检测0、10、25、50、100和150 μmol?L^-1 taurolidine诱导B16-F10细胞凋亡的百分比；细胞 克隆实验检测放射增敏性；Western blotting分析检测蛋白的表达。结果：50 μmol?L^-1 taurolidine作用于B16-F10细胞48 h，可诱导5%的细胞凋亡,随作用时间的延长及taurolidine浓度的增加，细胞凋亡的百分率增加（P<0.05）,呈显著的时效和量效关系；细胞克隆存活实验显示,当25 μmol?L^-1 taurolidine与2 Gy X射线照射联合作用B16-F10细胞时，与单纯给药组和单纯照射组比较，给药联合放疗组的细胞存活率显著降低（P<0.05），并随着药物浓度及照射剂量的增加细胞存活率进一步下降。随着taurolidine的浓度逐渐提高，小鼠黑色素瘤细胞放射增敏比（SER D₀和SER D_q）增高,50 μmol?L^-1 taurolidine组与10 μmol?L^-1组比较差异有显著性(P<0.05)；同时促凋亡蛋白Bax和Bad的表达增强，抗凋亡蛋白Bcl-2的表达下降。结论：taurolidine联合X射线照射通过调节Bcl-2家族蛋白协同诱导小鼠黑色素瘤细胞凋亡，从而提高了放射敏感性。

Effects of taurolidine on radiosensitivity of murine melanoma cells and its mechanism

Objective To observe the effects of taurolidine on radiosensitivity of B16-F10 cells of murine melanoma via the enhancement of Bax and Bad proteins and induction of Bcl-2 protein.Methods The apoptosis of B16-F10 cells was assessed after treated with 0,10,25,50,100 and 150 μmol·L-1 taurolidine,clone survival assay was used to detect the radiosensitivity of B16-F10 cells,and protein expressions were determined by Western blotting.Results The apoptosis of 5% cells was induced in a dose-and time-dependent manner after B16-F10 cells were treated with 50 μmol·L-1 taurolidine.The survival rate decreased after treated with taurolidine in combination with 2 Gy X-irradiation with the increase of taurolidine concentration and doses of irradiation(P<0.05),and the ratio of radiosensitivity (SER D0 and SER Dq) also increased with the increase of its concentration,there was significant difference between 50 μmol·L-1 taurolidine group and 10 μmol·L-1 taurolidine group(P<0.05); meantime,the level of proapototic protein Bax and Bad increased and the level of antiapoptotic protein Bcl-2 reduced.Conclusion Taurolidine in combination with irradiation can enhance the radiaosensitivity by the mediation of Bcl-2 family protein.