血根碱对人脑胶质瘤A172细胞生长及放射敏感性的影响

Effect of sanguinarine on growth and radiosensitivity of human glioma A172 cells

目的探讨血根碱对人脑胶质瘤A172细胞的生长及放射敏感性的影响。方法采用MTT法检测血根碱处理后A172细胞的生长和增殖；细胞流式技术测定细胞周期改变以及活性氧（ROS）的变化；磷脂酰丝氨酸外翻分析法检测细胞凋亡；细胞克隆形成法观察血根碱和辐射的联合作用。结果血根碱明显抑制脑胶质瘤A172细胞的生长，12、24和48h血根碱处理导致细胞生长抑制的半数抑制浓度（IC50）分别为4．83、3．91和3．25Ixmol／L。血根碱可以诱导剂量依赖性的细胞周期s期阻滞。与未处理的对照组相比，5Ixmol／L血根碱处理A172细胞24h后，早期凋亡细胞比例从（2．92±0．83）％增至（60．01±3．73）％，晚期凋亡细胞从（0．64．4-0．19）％增至（2．704-0．18）％，坏死细胞从（0．52±0．19）％增至（3．93±0．76）％，差异均具有统计学意义（t=55．28、8．32和9．51，P〈0．05）。另外，血根碱可以诱导ROS的产生，5μmol／L血根碱处理后ROS水平是对照组的4．1倍，而抗氧化剂氮乙酰半胱氨酸（NAC）能有效地抑制血根碱的细胞毒作用。采用多靶单击模型拟合曲线发现，与单纯照射组相比，血根碱预处理＋照射的联合处理组的放射增敏比（SERD0）达1．48。结论血根碱可以通过诱导细胞周期阻滞、细胞凋亡和ROS的产生抑制脑胶质瘤A172细胞的生长，而且在低剂量下，血根碱预处理可增加放射敏感性。

Objective To study the effect of sanguinarine on growth and radiosensitivity of human glioma A172 cells. Methods MTT assay was used to evaluate cell growth. Cell cycle analysis and reactive oxygen species（ROS） burst were performed by flow cytometry assay. Annexin V/PI assay was used to detect cell apoptosis. Colony formation assay was used to detect the influence of sanguinarine on cell radiosensitivity. Results Exposure of A172 cells to sanguinarine led to dose- and time- dependent eytotoxieity with IC50 values of 4.8, 3.9 and 3.2 μmol/L for 12, 24 and 48 h, respectively. Furthermore, sanguinarine caused an arrest of S phase. After being treated with 5 μmol/L sanguinarine for 24 h, the ratios of early apoptosis, late apoptosis and necrosis were （60. 01 ± 3.73 ） % , （2. 70 ± 0. 18 ） % and （3.93 ± O. 76）% respectively in A172 ceils compared with the untreated control（ t = 55.28, 8.32, 9.51 , P 〈 0. 05）. An increased generation of ROS was found after treatment with sanguinarine, however, NAC inhibited the effect of sanguinarine. As analyzed with multi-target click model fitting curves, the SERoo of sanguinarine-treated cells was 1.48. Conclusions Sanguinarine inhibits the A172 cells growth via apoptosis induction and ROS burst. Moreover, sanguinarine at a non-cytotoxicity dose increases eell sensitivity to X-rays.