目的 观察表皮生长因子受体单克隆抗体C225与60Co γ线对人肺鳞癌细胞系(H-520)的杀伤作用,探讨C225联合照射治疗非小细胞肺癌的可行性.方法 细胞成克隆实验中单纯照射(对照组)和照射+100 nmol/L C225组(实验组)给予0、1…2 4 6、8、10 Gy照射,培养10 d后计数≥50个细胞克隆.采用多靶单击模型进行数据处理,拟合细胞存活曲线,计算增敏比(D.值比).细胞凋亡实验均照射0、2、4、8 Gy后继续培养72 h,收集细胞,流式细胞仪检测细胞凋亡.细胞周期检测设对照组(不处理)、C225组(100 nmol/L)、照射组(8 Gy照射)和C225联合照射组(100 nmol/LC225+8 Gy),照后48 h收集细胞,流式细胞仪检测周期分布.结果 细胞克隆存活实验结果表明,扣除C225毒性影响后实验组存活分数比对照组低(F=6.36,P＜0.05),增敏比为1. 35.细胞凋亡实验结果显示,C225组4个剂量点凋亡百分率分别为13.75%±0.83%、25.12%±1.60%、46.12%.4-2.60%、50.51%±4.06%,对照组的分别为5.56%±0.62%、13.86%±0.80%、25.36%±1.02%、29.89%±2.09%(F=4.72,P＜0.05).细胞周期分布显示100 nmo//L C225可使细胞阻滞于G0+G1期,单纯照射阻滞于G2+M期,两者联合后同时出现G0+G1、G2+M期阻滞,三者均使S期细胞比例下降.结论 C225对H-520细胞具有放射增敏作用,其机制可能与细胞G0+G1期阻滞和诱发凋亡有关;结果 为临床上二者联合治疗非小细胞肺癌提供了理论基础.%Objective To investigate the efficacy of C225(cetuximab),a chimeric human-mouse anti-epithelial growth factor receptor monoclonal antibody.combined with 60Co gamma irradiation against humall non-small cell lung cancer cell line H-520. Methods H-520 cells were treated either with different dose of 60Co irradiation(1,2,4,6,8 and 10 Gy)alone or together with C225(100 nmol/L).Colony forming capacity was determined to create the survival curve 10 days after the treatment.Cells in different groups were harvested 72 hours after irradiation for apoptosis analysis or 48 hours after irradiation for cell cycle analysis by flow cytometry assay. Results The clone number in combinational treatment group was less than that in irradiation only group,which suggested that the cell survival rate in the combinational treatment group was significantly decreased comparing with irradiation only group(F=6.36,P＜0.05).The sensitizing enhance rate(SER)was 1.35.The percentage of apoptotic H-520 cells was 5.56%±0.62%,13.86%±0.80%,25.36%±1.02%and 29.89%±2.09%,respectively in 0,2,4 and 8 Gy irradiation alone groups,which were significantly lower than 13.75%±0.83%.25.12%±1.60%.46.12%±2.60%and 50.5l%±4.06%.respectively in irradiation combined with C225 treatment groups(F=4.72,P＜0.05).The cellcycle analysis showed that cells arrested in G0+G1 phases for C225 treatment,in G2+M phases for 60Co irradiation,and in both G0+G1 and G2+M phases for C225 in combination with 60Co irradiation. Conclusions C225 has radiosensitizing effects on H-520 cells.which may through the enhancement of 60Co irradiation-induced cell death and cell cycle arrest.This study provides a supportive evidence for clinical treatment in non-small cell lung cancer.
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