甘氨双唑钠对Tca-8113细胞体外放射增敏及黏附作用的影响

摘要

背景与目的:舌鳞状细胞癌是口腔癌中发病率较高的一种,治疗方法以手术加放疗为主.然而放疗常因射线对正常组织损伤或肿瘤对射线的抵抗而受到影响.因此,选择良好的放射增敏剂已成为优化舌癌治疗的新途径.本研究旨在探讨甘氨双唑钠(glycididazole sodium for injection,CMNa)联合射线对舌鳞状细胞癌Tca-8113细胞放射增敏及黏附的影响.方法:取对数生长期的Tca-8113细胞作为研究对象,MTT比色实验检测CMNa对Tca-8113细胞增殖的影响.克隆形成实验检测CMNa联合不同放射剂量对Tca-8113细胞克隆形成能力的影响,用多靶单击模型拟合细胞生存曲线,得出放射生物学敏感参数Do(平均致死剂量)、Dq(准阈剂量),并计算放射增敏比.流式细胞术FITC和PI双染比较不同放射剂量与CMNa联合不同放射剂量射线处理后细胞凋亡的变化.同质黏附实验检测CMNa联合放射处理前后细胞黏附能力的改变.结果:Tca-8113细胞的存活率随CMNa作用浓度及作用时间的延长而降低.克隆形成实验中求得单纯放射组的Do值为2.50,CMNa联合放射线处理的Do值为1.48,放射增敏比为1.68.流式细胞术检测CMNa联合不同放射剂量(2、4和6 Gy)的细胞凋亡率(49.6%、63.1%和78.8%)均显著高于单纯放射组(30.0%、53.9%和61.2%,P＜0.01).同质黏附实验在振荡20 min时,单纯放射组及CMNa+放射组与对照组相比,黏附细胞数显著增高(P＜0.01);但单纯放射组与CMNa+放射组相比黏附细胞数差异无统计学意义(P＞0.05).在振荡40和60 min时,单纯放射组及CMNa+放射组与对照组相比,黏附细胞数显著增高(P＜0.01);且(CMNa+放射组与单纯放射组相比,黏附细胞数差异有统计学意义(P＜0.01).结论:CMNa对Tca-8113细胞有明显的放射增敏作用,其机制可能与促进细胞凋亡有关,并可能改变Tca-8113细胞的同质黏附能力.%Background and purpose: Tongue squamous cell carcinoma is a high incidence disease in oral cancers. Surgery and radiotherapy are the main methods for the treatment for tongue cancer. However, the effect of radiotherapy is limited owing to injured healthy tissues and the resistance of tumor tissues. This study aimed to investigate radiosensitivity and homogeneous adhesion of CMNa with different doses of radiotherapy to human tongue cancer Tca-8113 cell line. Methods: Tca-8113 cells in the logarithmic growth phase were used for experimentation.Cell growth inhibition was assessed using MTT after being treated with various concentrations and durations of time.The mean lethal dose (Do) and sensitive enhancement ratio (SER) were obtained through colony formation assay after different treatments. FITC and PI, double labels, detect apoptosis in order to compare the difference between radiotherapy and CMNa with different doses of radiotherapies by flow cytometry. The changes in the cell homogeneous adhering capacity were observed by homotypic adhesion experiment. Results: Surviaval rates of Tea-8113 cells were decreased as CMNa increased in concentration and time. Apoptosis rates of the combination groups (49.6％, 63.1％and 78.8％) were significantly higher than in the radiotherapy groups (30.0％, 53.9％ and 61.2％, P＜0.01). The Do and SER of the combination groups were 1.48 and 1.68. The Do of radiotherapy only was 2.50. The homotypic adhesion experiments showed that there were significant differences in the numbers of adhering cells among all the 3 groups after 40 and 60 minutes of vibration (P＜0.01). Conclusion: CMNa can significantly enhance the radiosensitivity of human tongue cancer Tca-8113 cells with different doses of irradiation. The mechanism could be related to enhanced apoptosis.CMNa also could change homogeneous adhering capacity.