奥沙利铂对人肝癌细胞HepG2放射增敏作用初探  

The effect of oxaplatin on enhancing radiosensitivity in human hepatocellular carcinoma cell line HepG2

作　　者：王健仰[1] 杨伟志[1] 冯燕茹[1] 肖琴[1] 任骅[1] 金晶[1] 李晔雄[1] Wang Jianyang Yang Weizhi Feng Yanru Xiao Qin Ren Hua Jin Jing Li Yexiong(Department of Radiation Oncology , Cancer Hospital （ Institute）, Chinese Academy of Medical Sciences, Peking Union Medical College ,Beiiing 100021, China)

机构地区：[1]国家癌症中心、中国医学科学院北京协和医学院肿瘤医院放疗科,北京100021

出　　处：《中华放射肿瘤学杂志》2016年第11期1255-1260,共6页Chinese Journal of Radiation Oncology

基　　金：国家重大研发计划项目（2016YFC0904600）

摘　　要：目的：观察奥沙利铂对人肝癌细胞HepG2放射后增殖的影响。方法采用MTT法测定不同浓度奥沙利铂作用HepG2细胞不同时间后的细胞半数抑制浓度（ IC50）。选择合适的奥沙利铂浓度行细胞克隆形成实验，分别给予3种不同处理：HepG2空白组、单纯照射组（1、2、4、6、8、10 Gy单次照射）、奥沙利铂＋照射组）。同期选取6 Gy照射组检测细胞凋亡情况。检测以上3组细胞中细胞增殖蛋白ERK 1／2和DNA损伤修复蛋白Ku.70表达情况。结果奥沙利铂对HepG2细胞的6、12、24、48 h的IC50分别为54.4、29.1、17.8、10.5 mg／L。3 mg／L浓度奥沙利铂的放射增敏比为1.59。细胞凋亡情况检测结果表明奥沙利铂＋照射组存活细胞比例均低于照射组（ P＝0.005）、奥沙利铂组（P＝0.008）和空白对照组（P＝0.001）。照射组和奥沙利铂＋照射组的ERK 1／2表达水平被持续抑制，其中照射组的ERK 1／2表达水平在处理后48 h开始升高，但低于对照组，奥沙利铂＋照射组的ERK 1／2表达水平一直为最低，且没有升高趋势。各组Ku.70的变化水平与ERK 1／2类似。结论奥沙利铂对于人肝癌细胞HepG2的体外放射具有显著的增敏作用。Objective To explore the effect of oxaliplatin （ OXA） on enhancing radiosensitivity in human hepatocellular carcinoma cell line HepG2 . Methods 50% inhibition concentration （ IC50 ） of HepG2 cells treated with OXA was measured by using MTT method at 6, 12, 24, 48 hours. Then clone formation assay was applied to obtain sensitizing enhancement ratio （ SER） of OXA combing IR, according to the survival fraction of three groups 10.14 days after treatments：placebo.treated group （ C） ,radiation group （ IR, single dose of 1 Gy,2 Gy,4 Gy,6 Gy,8 Gy,10 Gy） and IR synchronizing OXA group （ IR＋3 mg/L OXA） . The proportions of cell apoptosis were analyzed using flow cytometry at 24 hours after treatment. At last, we semi.quantitative tested the expression of extracellular regulated protein kinase 1/2 （ ERK 1/2 ） and DNA damage repair protein Ku.70 of the C,IR and IR＋OXA groups. Statistical analysis was performed by T test. Results The IC50 of OXA on HepG2 cells is 54.4 mg/L at 6 hours,29.1 mg/L at 12 hours,17.8 mg/L at 24 hours and 10.5 mg/L at 48 hours.3 mg/L was selected in clone formation assay at which 80.90% HepG2 cells survived at 24 hours. The SER （ SF2 ） is calculated as 1.59. Flow cytometry showed the proportion of survival cells in IR＋OXA group is significantly lower than those of IR group （ P=0.005） ,OXA group （ P=0.008） and C group （ P=0.001） . The expressions of ERK 1/2 were inhibited in IR and IR＋OXA groups compared by that of control group. But the expression of ERK 1/2 in IR group showed increasing after 48 hours which was higher than that of IR＋OXA group. For Ku.70,the changes of expression were similar with that of ERK 1/2. Conclusion Oxaliplatin presented enhancing radiosensitivity in human hepatocellular carcinoma cell line HepG2 in vitro.

关 键 词：细胞系 肝癌 奥沙利铂 放射增敏比 

分 类 号：R735.7[医药卫生—肿瘤]