摘要
目的探讨三氧化二砷(As2O3)联合热疗对食管癌细胞株EC-1放疗的增敏效果,并探讨其可能机制。方法将人食管癌细胞株EC-1分为4组,分别为单纯放疗组(R组)、热疗+放疗组(HR组)、As2O3+放疗组(AR组)、As2O3+热疗+放疗组(AHR组),采用平板克隆形成实验检测细胞放射敏感性,通过单击多靶模型拟合细胞存活曲线。采用流式细胞仪检测细胞周期及细胞凋亡率。所有实验均重复3次。结果平板克隆形成实验显示:R组、HR组、AR组、AHR组平均致死剂量(D0)依次为2.192、2.070、1.705、1.272 Gy;准阈剂量(Dq)依次为0.461、0.241、0.134、0.042 Gy;靶点数(N)依次为1.625、1.308、1.192、1.079;HR组、AR组、AHR组平均致死剂量增敏比(SERD0)依次为1.059、1.286、1.723,准阈剂量增敏比(SERDq)依次为1.917、3.474、11.000。流式细胞术检测细胞周期,发现HR组、AR组及AHR组细胞G0/G1期、S期比例较R组降低,G2/M期比例较R组增加(P<0.05);AR组细胞G0/G1期比例较HR组降低(P<0.05);AHR组细胞各周期比例与HR组、AR组比较均有差异(P<0.05)。HR组、AR组及AHR组细胞凋亡率较R组升高(P<0.05);AHR组细胞凋亡率亦高于HR组、AR组(P<0.05)。结论 As2O3和热疗均可以增加食管癌细胞株EC-1对放疗的敏感性,二者联合应用增敏作用更明显。可能机制是使细胞阻滞在G2/M期,使G0/G1期、S期细胞比例减少;其次是通过诱导细胞凋亡,增强放射线对细胞的杀伤作用。
Objective To investigate the radiosensitizing effect and underlying mechanism of arsenic trioxide(As2O3) in combination with hyperthermia on human esophageal squamous carcinoma cell line EC-1. Methods There were four groups as the irradiation group( R),the hyperthermia with irradiation group( HR),the As2O3 with irradiation group(AR) and the As2O3 with hyperthermia and irradiation group(AHR). Clonogenic radiation survival assays on EC- 1 cell line after treatments with As2O3 and hyperthermia with or without radiation were adopted. Cell survival curve was gained using single-hit multitarget model. Cell cycles and the cell apoptosis rate of the 4 different groups were tested by flow cytometric analysis(FCM). All experiments were repeated 3 times. Results According to the clonogenic radiation survival assay,the radiobiological parameters D0,Dqand N in the R,HR,AR and AHR groups was 2. 192 Gy,2. 070 Gy,1. 705 Gy,1. 272 Gy; 0. 461 Gy,0. 241 Gy,0. 134 Gy,0. 042 Gy; 1. 625,1. 308,1. 192,1. 079. The SERD0 and SERDqin the HR,AR and AHR groups was 1. 059,1. 286,1. 723; 1. 917,3. 474,11. 000. The FCM showed that cells in G0/G1 phases and S phases were significantly lower in HR,AR and AHR groups than R group(P < 0. 05),while cells in G2/M phase were significantly higher in HR,AR and AHR groups than R group(P < 0. 05). Cells in G0/G1 phases were lower in AR group than HR group( P <0. 05). Cell cycles in AHR group was obvious different compared to AR and HR groups( P < 0. 05). Cell apoptosis was enhanced in other 3 groups compared to R group(P < 0. 05). Among the 3 groups,cell apoptosis in AHR group was higher than HR and AR groups(P < 0. 05). Conclusion Both As2O3 and hyperthermia can enhance the radiosensitivity in EC- 1,and combination of them could enhance this effect more significantly. The change of EC- 1 cell cycle,including the increase of G2/M and decrease of G0/G1 and S,may be responsible for the radiosensitivity effect. Meanwhile,the induction of apoptosis which is benefit to the killing effect of radiotherapy on EC- l cell was also involved in this process.
引文
1 张佃富,张培良,王玉国,等.食管癌全程三维适形放疗后程加速超分割的疗效观察[J].中华肿瘤防治杂志,2010,17(7):521-522,532.
2 邱嵘,王军,王玉祥,等.三维适形与调强放疗治疗食管癌的临床对照研究[J].中华肿瘤防治杂志,2012,19(1):68-71.
3 王坚,卢绪箐,李毅,等.热疗对放射性肺损伤保护作用的实验研究[J].中华肿瘤防治杂志,2010,17(21):1714-1717.
4 杨绍灵,凌如芳,许志金.全反式维甲酸、三氧化二砷联合化疗治疗初治急性早幼粒细胞白血病的临床观察[J].疑难病杂志,2010,9(3):194.
5 宿颖,杨慧,张敏,等.三氧化二砷联合放疗对口腔癌移植瘤小鼠治疗作用的研究[J].口腔生物医学,2010,1(4):173-177.
6 郭红,于洪,王天禄.三氧化二砷对人肺癌A549细胞的放射增敏作用[J].中国肿瘤,2012,21(10):784-786.
7 王维虎,杨伟志,李晔雄.三氧化二砷对纤维肉瘤细胞放射增敏作用的研究[J].中华放射肿瘤学杂志,2006,15(6):448-451.
8 黄辉,蒋宏玲,刘芬菊,等.60Coγ射线照射联合三氧化二砷对人脑胶质瘤SHG44细胞DNA损伤的研究[J].现代中西医结合杂志,2012,21(12):1274-1276.
9 余桂芳,严跃红,陈益民.三氧化二砷对人肝癌细胞生长及其侵袭转移能力的影响[J].疑难病杂志,2011,10(4):293.
10 孙秉赋,张博,栾明翰.三氧化二砷联合氟尿嘧啶对人肝癌SMMC-7721细胞Bcl-2和Bax表达的影响[J].中国煤炭工业医学杂志,2013,16(11):1861-1863.
11 景绍武,王雅立,吴风鹏,等.三氧化二砷与放射线联合对食管癌细胞株Eca109的增敏作用及机制[J].肿瘤防治研究,2011,38(6):620-623.
12 唐知己,章文斌,张锐,等.125I近距离放疗联合热疗对人胶质瘤细胞毒性的研究[J].临床神经外科杂,2010,7(1):14-16.
13 董军兰,姚洁.同步放化疗联合深部热疗治疗中晚期宫颈癌的临床疗效观察[J].生物医学工程进展,2011,32(4):233-235.
14 马姗姗,王仁生,肖帅,等.鼻咽癌热疗疗效的系统评价[J].中华肿瘤防治杂志,2011,18(4):288-291.
15 谌龙,孙娟,马春峰,等.非小细胞肺癌放疗联合体外高频热疗80 例疗效观察[J].中国现代药物应用,2012,6(18):46-47.
16 张佳慧,秦丽娟.肿瘤热疗的研究进展[J].实用心脑肺血管病杂志,2012,20(9):1424.
17 郭可锋,柳玉华,李宗民.三维适形放疗联合射频热疗治疗中晚期食管癌的疗效观察[J].实用癌症杂志,2011,26(1):67-77.
18 李征,米登海,杨克虎,等.放疗联合热疗治疗食管癌的Meta分析[J].中华肿瘤防治杂志,2012,19(9):671-675.
19 谌龙,马春峰,孙娟,等.食管癌放疗联合体外高频热疗40例疗效观察[J].中国煤炭工业医学杂志,2012,15(9):1348-1349.
20 梁克.肿瘤分子生物放射生物学研究进展[M]//殷蔚伯.肿瘤放射治疗学.3版.北京:北京医科大学中国协和医科大学联合出版社,2002:430-432.
21 李岚,张敬东,刘云鹏,等.三氧化二砷对胃癌BGC-823细胞增殖及细胞周期的影响[J].肿瘤学杂志,2008,14(3):190-192.
22 Park WH,Seol JG,Kim ES,et al.Arsenic trioxide-mediated growth inhibition in MC/Car myeloma cells via cell cycle arrest in association with induction of cyclin-dependent kinase inhibitor,p21,and apoptosis[J].Cancer Res,2000,60(11):3065-3071.
23 刘敩,夏轶男,张晖.三氧化二砷联合热疗Hep-2细胞周期进程及凋亡率的影响[J].老年医学杂志,2012,32(4):753-754.
