青蒿琥酯对人宫颈癌HeLa细胞的放射增敏作用

Study of radiation sensitization of artesunate on human HeLa cells of cervical cancer

目的 探讨表蒿琥酯对人宫颈癌HeLa细胞的放射增敏作用.方法 应用60Cor射线照射细胞,吸收剂量率为0.635 Gy/min,照射剂量为0、1、2、4和6 Gy.采用MTT法检测青蒿琥酯对HeLa细胞的抑制作用,确定青蒿琥酯的最适实验作用浓度.克隆形成法检测青蒿琥酯对HeLa细胞放射敏感性的影响;采用＂多靶单击数学模型＂拟合HeLa细胞的剂量-存活曲线,得出平均致死剂量、准阈剂量及放射增敏比,评价其增敏效果.用流式细胞术检测HeLa细胞凋亡率,进一步检测青蒿琥酯对HeLa细胞的放射增敏性.结果 青蒿琥酯对HeLa细胞的抑制作用随药物浓度的增加而增加,单纯照射1、2、4和6 Gy细胞的克隆形成率分别为91.67%、82.02%、58.60%和25.01%,加入青蒿琥酯后,相同照射剂量下克隆形成率分别降低为74.93%、60.53%、22.38%和5.05%;单纯照射组与药物＋照射组的平均致死剂量（D0）分别为2.95和2.07 Gy,准阈剂量（Dq）分别为2.01和1.24 Gy,放射增敏比（SER）为1.43.单纯2和6 Gy照射组细胞凋亡率分别是12.26%和40.08%,加入青蒿琥酯后,相同照射剂量下细胞凋亡率分别上升至22.71%和59.92%.结论 青蒿琥酯对人宫颈癌HeLa细胞的抑制作用成药物浓度依赖性;青蒿琥酯对HeLa细胞具有一定的放射增敏作用.

Objective To investigate the radiosensitizing effects of artesunate on human HeLa cells of cervical cancer in vitro.Methods Hela cells were irradiated with 60Co γ-rays.The dose rate was 0.635 Gy/min and the radiation dose was 0,1,2,4,6 Gy,respectively.The anti-proliferation activities of artesunate on HeLa cells were evaluated with MTT assay,to determine the most appropriate drug concentration.The effect of radiosensitivity was observed by using clonogenic assay.The single-hit multitarget model was used to plot the HeLa cell＇s dose-survival curve,to calculate mean lethal dose,quasithreshold dose and sensitization enhancement rate,and to evaluate its radiosensitization effect.The apoptosis was analyzed with flow cytometry （FCM） to further test the radiation senseitization of artesunate on HeLa cells.Results The inhibition of artesunate on HeLa cells increased with concentration.In radiation group,the cell cloning efficiency were 91.67% ,82.02% ,58.60% ,25.01%,respectively,and in artesunate （2.0 μ mol/L） ＋ radiation group,the cell cloning efficiency were 74.93% ,60.53% ,22.38% ,5.05%.In radiation group and artesunate （2.0 μmol/L） ＋ radiation group,the mean lethal dose（D0） was 2.95 and 2.07 Gy,respectively,while the qusai-threshold dose （Dq） were 2.01 and 1.24 Gy,respectively,and SER was 1.43.Compared with 2 and 6 Gy radiation group,the apoptosis rate of drug ＋ radiation group increased from 12.26% ,40.08% to 22.71% ,59.92%.Conclusions The inhibiting effect of artesunate on HeLa cells is concentration-dependent.Artesunate has radiosensitizing effect on HeLa cells in vitro.