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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IC50(95%CI)。在常氧和缺氧条件下,分别给予0、1、2 Gy单次吸收剂量照射经1 mmol/L CMNa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,Western blotting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G1期比例升高,S期比例降低;促凋亡蛋白Caspase 3/8/9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Cyclin D1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 effect of CMNa on the radiosensitivity of human neuroblastoma cell line SH-SY5Y under normoxia and hypoxia conditions. Methods SH-SY5Y cells were treated with different concentrations of CMNa under normoxia and anaerobic conditions. The IC50 value and the 95% confidence interval (IC50) were determined by CCK-8 assay when the cell survival rate was 50% ). Under normoxia and hypoxia conditions, SH-SY5Y (CMNa group) and SH-SY5Y without CMNa treatment (control group) were treated with 1 mmol / L CMNa, The formation of colony was observed. The changes of cell cycle and apoptosis were detected by flow cytometry. The expression of cell cycle and apoptosis related proteins were analyzed by Western blotting. Results IC50 (95% CI) of SH-SY5Y were 3.62 (3.07-4.26) and 1.59 (1.20-2.12) mmol / L, respectively, under normoxia and hypoxia conditions. Under normoxia, the number of colony formation in CMNa group was significantly less than that in control group. Compared with normoxia state, the number of clonal formation in hypoxia decreased significantly. Under the condition of hypoxia, compared with the control group, the apoptosis rate of cells in CMNa group and the proportion of cells in G1 phase increased and the proportion of S phase decreased; the expression of Caspase 3/8/9 and Bax increased, and the expression of Bcl- -2 expression decreased; cell cycle related protein Cyclin D1 expression decreased p16 expression increased. Conclusion CMNa has a specific radiosensitizing effect on SH-SY5Y in hypoxia.