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目的研究重组人血管内皮抑制素(恩度)改善乏氧、增强肺腺癌细胞系A549的放射敏感性效果。方法①在常氧和乏氧培养下,以不同浓度的恩度与A549细胞分别处理不同的时间,用MTT法分别选择生长抑制率为20%的药物浓度(IC20);②将A549细胞分别在常氧和乏氧培养下分为对照组和恩度组,加药培养24 h后,分别给予高能X线0、2、4、6、8 Gy剂量照射。集落形成率实验检测各组在不同条件下的细胞存活分数,根据多靶单击模型拟合细胞存活曲线,求出各组的平均致死剂量(D0)、准域剂量(Dq)、外推数N、照射2 Gy时的细胞存活率(SF2)、放射增敏比(SER);③将A549细胞分别在常氧和乏氧培养下,分成对照组、单放组、恩度组和联合组。4组分别处理24 h后,通过流式细胞仪对各组凋亡细胞进行定量检测,得出各组的凋亡率。结果①恩度可以抑制常氧和乏氧培养下的A549细胞的生长,且呈时间-剂量依赖性,IC20分别是(330.34±4.18)mg/L和(437.23±3.32)mg/L;②在多靶单击模型中,在常氧下对照组和恩度组的D0值分别是1.36、1.30,Dq值分别是1.02、1.02;在乏氧培养下对照组和恩度组的D0值分别是1.70、1.39,Dq值分别是2.45、1.03。常氧和乏氧组恩度的SER分别为1.04和1.22;③在常氧培养下,对照组、单放组、恩度组和联合组的凋亡率分别为(15.91±0.57)%、(42.70±0.37)%、(19.90±0.48)%、(41.52±0.38)%,联合组和单放组比较,差异无统计学意义(F=1.22,P=0.479);在乏氧培养下4组的凋亡率分别为(16.72±0.67)%、(30.11±0.95)%、(26.71±0.62)%、(36.34±0.71)%,联合组和单放组比较,差异有统计学意义(F=3.34,P=0.036)。结论恩度在常氧培养下没有放射增敏作用,细胞凋亡率没有明显增加;而在乏氧培养下能提高A549的放射敏感性,而且细胞凋亡率明显增加。
Objective To study the effect of recombinant human endostatin (Endostar) on hypoxia and enhance the radiosensitivity of lung adenocarcinoma cell line A549. Methods ① Under normoxia and hypoxia culture, A549 cells were treated with A549 cells at different concentrations for different time. MTT assay was used to select the drug concentration with 20% inhibition (IC20) Under normoxia and hypoxia culture, the rats were divided into control group and Endu group. After dosing for 24 hours, they were given high-energy X-rays 0, 2, 4, 6 and 8 Gy respectively. The colony formation rate was used to test the cell survival score of each group under different conditions. The survival curves of cells were fitted according to multi-target click model, and the average lethal dose (D0), quasi-domain dose (Dq), extrapolation number N, the cell survival rate (SF2) and the radiosensitization ratio (SER) when exposed to 2 Gy; ③The A549 cells were divided into control group, single release group, Endostar group and combination group under normoxia and hypoxia culture respectively, . 4 groups were treated for 24 h, by flow cytometry quantitative detection of apoptotic cells in each group, the rate of apoptosis was obtained in each group. Results Endostar could inhibit the growth of A549 cells under normoxia and hypoxia culture in a time-and dose-dependent manner with (330.34 ± 4.18) mg / L and (437.23 ± 3.32) mg / L, respectively. In the multi-target click model, the D0 values of control group and Endu group under normoxia were 1.36 and 1.30, respectively, and the Dq values were 1.02 and 1.02 respectively. The values of D0 of control group and Endu group under hypoxic culture were 1.70,1.39, Dq values were 2.45,1.03. The SERs of normoxia and hypoxia groups were 1.04 and 1.22, respectively. ③The apoptosis rates of control group, radiotherapy group, endostar group and combination group were (15.91 ± 0.57)% and (F = 1.22, P = 0.479). There was no significant difference between the combined group and the single group (P> 0.05) (16.72 ± 0.67)%, (30.11 ± 0.95)%, (26.71 ± 0.62)%, (36.34 ± 0.71)%, respectively. The difference between the combined group and the single group was statistically significant (F = 3.34, P = 0.036). Conclusion Endostar had no radiosensitization effect under normoxia culture and no significant increase of apoptosis rate. Under hypoxia culture, it increased the radiosensitivity of A549 and markedly increased the apoptosis rate.