摘要
目的探讨甘氨双唑钠（CMNa）在常氧和缺氧条件下对人神经母细胞瘤细胞株（SH—SY5Y）放射敏感性的影响。方法在常氧和缺氧条件下，以不同浓度的CMNa处理SH—SY5Y，CCK-8法检测细胞存活率为50％时的药物浓度（IC50）及其95％可信区间Ic，。（95％CI）。在常氧和缺氧条件下，分别给予0、1、2Gy单次吸收剂量照射经1mmol／LCMNa处理的SH—SY5Y（CMNa组）和未经CMNa处理的SH—SY5Y（对照组），观察克隆形成情况，流式细胞术检测细胞周期变化和细胞凋亡率，Westernblotting分析细胞周期和凋亡相关蛋白的表达变化。结果常氧和缺氧条件下，SH·SY5Y的IC50（95％CI）分别为3．62（3．07—4．26）和1．59（1．20—2．12）mmol／L。在常氧状态下，CMNa组克隆形成数量显著少于对照组；与常氧状态比较，缺氧状态下的克隆形成数量明显减少。在缺氧条件下，与对照组比较，CMNa组细胞凋亡率和细胞G，期比例升高，S期比例降低；促凋亡蛋白Caspase3／8／9和Bax表达增加，抑制凋亡蛋白Bcl-2表达减少；细胞周期相关蛋白CyclinD1表达减少，p16表达增加。结论CMNa对缺氧状态下的SH—SY5Y有特异的放疗增敏作用。
Objective To investigate the effects of glycidiazale sodium （CMNa） on radiosensitivity of children＇s neuroblastoma SH-SY5Y cell line under normoxia and hypoxia conditions. Methods CMNa in different concentration was used on SH-SY5Y under normoxia and hypoxia. CCK-8 was employed to determine half maximal （50%） inhibitory concentration （IC） and 95% confidence interval. Radiation absorbed doses of 0, l, and 2 Gy were given to SH-SY5Y with 1 mmol/L CMNa （CMNa group） and normal SH-SYSY （control group）. Colony-forming assay was conducted to explore the cell proliferation. Flow cytometry was conducted to test cell cycle and their apoptosis. Western blotting was employed to test expression changes of cell cycle and apoptosis-related proteins. Results Under normoxia and hypoxia conditions, values of IC50（95% CI） of SH-SY5Y were 3.62 （3.07 -4.26） mmol/L and 1.59 （ 1.20 - 2. 12） mmol/L, respectively. Under normoxia condition, clone formation quantity was significantly lower in CMNa group than that of control group. Compared to normoxia condition, clone formation quantity was significantly lower in hypoxia condition. Under hypoxia condition, compared to control group, cell apoptosis rate and Gl-phase ratio increased significantly; S-phase ratio decreased significantly; expressions of stimulated apoptosis protein （Caspase 3/8/9） and Bax increased; inhibited apoptosis protein （Be1-2） decreased; expression of cell cycle-related protein （Cyelin D1） decreased; and expression of P16 increased in CMNa group. Conclusion CMNa has specific enhancing effect on radiosensitivity of SH-SY5Y under hypoxia condition.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2014年第1期17-22,共6页
Journal of Shanghai Jiao tong University:Medical Science
基金
上海市市级医院新兴前沿技术联合攻关项目(SHDC12012110)
卫生部医药卫生科技发展研究中心项目(W2012FZ053)~~
