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在前期实验证实吴茱萸碱(Evodiamine,EVO)对舌鳞状细胞癌Tca8113细胞有放射增敏的作用,为进一步研究EVO作为放射增敏剂应用的可能性,并探讨不同加药时序对放射增敏作用的影响是否有差异,采用EVO联合射线作用于胃癌SGC7901细胞探讨EVO对其放射敏感性的影响。采用MTT法检测不同时间点、浓度EVO对SGC7901细胞增殖的抑制率,并检测低浓度EVO不同加药顺序联合射线后对SGC7901细胞增殖的抑制率;克隆形成实验分析联合作用对SGC7901细胞克隆形成能力的影响,拟合存活曲线并计算各组细胞放射增敏比;流式细胞术分析EVO联合放射线作用后细胞周期分布变化;Western blot检测凋亡相关蛋白caspase3的表达水平。通过上述实验方法得到EVO对SGC7901细胞的生长抑制作用呈时间、剂量依赖,并且不同加药顺序对细胞增殖的抑制效果不同(P<0.05)。克隆形成实验中求得联合作用组各放射剂量下的存活分数、放射生物学敏感参数均低于单独放射组,且联合作用组中先加药组的增敏效果优于后加药组(P<0.05)。流式细胞术检测EVO联合不同放射剂量使细胞周期阻滞在G2/M期的比率明显高于单独放射组及单独加药组(P<0.05)。Western blot结果提示联合作用组中凋亡蛋白caspase3的表达高于单独放射组及单独加药组(P<0.05)。综上所述,EVO对SGC7901细胞有放射增敏作用,并且不同加药顺序对放射增敏作用的影响不同,联合作用中先加药后放射的效果优于先放射后加药组,其增敏效果的作用机制可能与联合作用导致细胞G2/M期阻滞、凋亡蛋白caspase3表达增加有关。
Previous studies demonstrated that Evodiamine (EVO) has a radiosensitizing effect on tongue squamous cell carcinoma Tca8113 cells. In order to further investigate the potential of EVO as a radiosensitizer and to investigate the effect of different dosing schedules on radiosensitization The effects of EVO on the radiosensitivity of SGC7901 cells were evaluated using EVO combined with radiotherapy. The inhibitory rate of proliferation of SGC7901 cells at different time points and concentrations of EVO was detected by MTT assay. The inhibitory rate of SGC7901 cells proliferation was detected by different dosages of EVO combined with low concentration of EVO. Clonal formation assay was used to evaluate the clonogenic capacity of SGC7901 cells The survival curves were fitted and the radiosensitization ratio was calculated. The cell cycle distribution was analyzed by flow cytometry (FCM) and the expression of caspase3 was detected by Western blot. The growth inhibition effect of EVO on SGC7901 cells was time-and dose-dependent by the above experimental method, and the effect of different drug-adding sequences on cell proliferation was different (P <0.05). Survival scores and radiosensitivity parameters of radiosensitization in combination group were lower than that of radiotherapy alone group in colony formation experiment, and sensitization effect of combination group was superior to that of group D <0.05). The ratio of cell cycle arrest in G2 / M phase by flow cytometry in EVO combined with different radiation dose was significantly higher than those in radiotherapy alone group and drug alone group (P <0.05). Western blot results suggest that the caspase3 expression in the combined group is higher than that in the radiotherapy alone group and the single drug addition group (P <0.05). In summary, EVO had a radiosensitizing effect on SGC7901 cells, and the effect of different drug-loading sequences on radiosensitization was different. The combined effect of the first dosing and the second dosing was better than that of the first dosing and the second dosing Sensitive effect of the mechanism may be combined with the joint G2 / M phase lead to cell arrest, the expression of apoptosis protein caspase3 increased.