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目的探讨吴茱萸碱(Evo)对Eca-109食管鳞癌细胞放射敏感性的影响,探索其可能机制。方法采用MTT法检测(10、20、40、60、80、100、120)μg/m L吴茱萸碱对Eca-109细胞生长的抑制作用,确定最佳处理剂量。实验分为放射组(仅给予0、2、4、6、8 Gy X线照射处理)、联合组(给予80μg/m L吴茱萸碱和0、2、4、6、8 Gy X线照射处理),克隆形成实验检测吴茱萸碱对Eca-109细胞克隆形成能力的影响,流式细胞术检测细胞周期;Western blot法检测Eca-109细胞中Ku70、Ku80、DNA激活蛋白激酶催化亚基(DNA-PKcs)、DNA双链修复蛋白Rad51蛋白水平。结果吴茱萸碱呈剂量依赖性抑制Eca-109细胞的生长,80μg/m L吴茱萸碱的抑制作用最大。克隆形成实验结果显示80μg/m L吴茱萸碱联合放射治疗后,平均致死剂量(D0)、准域剂量(Dq)值显著小于单纯放射时的值,放射增敏比(SER)为1.86±0.06,拟合的生存曲线左移。吴茱萸碱可减少放射诱导的细胞G2/M期阻滞,可显著抑制放射诱导的Ku70、Ku80、DNA-PKcs、Rad51蛋白上调。结论吴茱萸碱具有增加食管鳞癌细胞株Eca-109放射敏感性的作用,可能与抑制细胞周期G2/M期阻滞和降低DNA损伤修复蛋白表达水平有关。
Objective To investigate the effect of evodiamine on radiosensitivity of esophageal squamous carcinoma cell line Eca-109 and to explore its possible mechanism. Methods The inhibitory effect of (10, 20, 40, 60, 80, 100, 120) μg / mL of evodiamine on the growth of Eca-109 cells was determined by MTT assay. The experiment was divided into radiation group (given 0, 2, 4, 6, 8 Gy X-ray treatment only), the combination group (80 μg / m L evodiamine and 0, 2, , The effect of evodiamine on clonality of Eca-109 cells was detected by clonogenic assay and the cell cycle was detected by flow cytometry. The expressions of Ku70, Ku80, DNA-PKcs catalytic subunit (DNA-PKcs ), DNA double-strand repair protein Rad51 protein level. Results Evodiamine inhibited the growth of Eca-109 cells in a dose-dependent manner with the inhibition of evodiamine at 80 μg / mL. The results of colony formation assay showed that the mean lethal dose (D0) and quasi-domain dose (Dq) of 80 μg / mL evodiamine combined with radiotherapy were significantly lower than those of pure radiotherapy. The radiosensitization ratio (SER) was 1.86 ± 0.06, Fitting the survival curve to the left. Evodiamine reduced the G2 / M phase arrest induced by radiation and significantly inhibited the up-regulation of radiation-induced Ku70, Ku80, DNA-PKcs and Rad51 proteins. Conclusions Evodiamine can increase the radiosensitivity of esophageal squamous carcinoma cell line Eca-109, which may be related to the inhibition of cell cycle G2 / M arrest and DNA damage repair protein expression.