尼妥珠单抗对食管鳞癌ECA-109和TE-13细胞放射敏感性的影响及其发生机制  

Effects and its mechanism of Nimotuzumab on radiosensitivity of esophageal carcinoma ECA-109 and TE-13 cell lines

作　　者：王军[1] 王雯[2] 郭银[2] 景绍武[2] 尚凯[2] 苗明昌[2] 王京[2] 武亚晶 刘丽娜[2] 于金明[3] Wang J Wang W Guo Y Jing SW Shang K Miao MC Wang J Wu YJ Liu LN Yu JM(Department of Radiation Oneology, Tianjin Medical University Cancer Institute ＆ Hospital, Tianjin 300070, China Department of Radiation Oncology, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China Department of Radiation Oncology, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China Department of Radiation Oncology, Shandong Cancer Hospital,Jinan 250117, China)

机构地区：[1]天津医科大学肿瘤医院放疗科,300070 [2]河北医科大学第四医院放疗科,石家庄050011 [3]山东省肿瘤医院放疗科,济南250117

出　　处：《中华肿瘤杂志》2016年第10期732-738,共7页Chinese Journal of Oncology

摘　　要：目的探讨尼妥珠单抗对食管癌ECA-109和TE-13细胞放射敏感性的影响及其发生机制。方法体外培养人食管癌ECA-109和TE-13细胞，分为空白组、照射组、药物组和联合组（照射＋药物治疗）。联合组ECA-109、TE-13细胞在照射前24h给予尼妥珠单抗处理，照射后2h收集细胞。克隆形成实验检测尼妥珠单抗对人食管癌ECA．109和TE-13细胞的放射增敏作用。流式细胞术检测各组细胞的凋亡情况。Westernblot检测表皮生长因子受体（EGFR）、磷酸化表皮生长因子受体（p-EGFR）、DNA依赖蛋白激酶催化亚单位（DNA-PKcs）、磷酸化DNA依赖蛋白激酶催化亚单位（p-DNA．PKcs）和磷酸化组蛋白H2AX（7H2AX）的表达。结果ECA-109细胞联合组的准阈剂量（Dq）、平均致死量（D0）和细胞受到2Gy照射后的存活分数（SF2）分别为1．11、1．40和0．42，照射组分别为1．72、2．14和0．66，差异均有统计学意义（均P〈0．05）；TE．13细胞联合组的Dq、D0和细胞受到2Gy照射后的SR分别为0．41、0．43和0．40，照射组为0．46、0．65和0．71，差异均有统计学意义（均P〈0．05）。尼妥珠单抗对ECA-109细胞和TE-13细胞的放射增敏比为1．35和1．43。流式细胞仪检测结果显示，ECA-109细胞和TE-13细胞联合组的细胞凋亡率均高于照射组，差异均有统计学意义（均P〈0．05）。Westernblot结果显示，ECA．109细胞和TE．13细胞各组的EGFR和DNA-PKcs的蛋白表达水平均无明显变化（均P〉0．05）。ECA-109细胞和TE，13细胞中，照射组p-EGFR和p-DNA．PKcs的表达水平均高于空白组，照射组和药物组γH2AX的表达水平均高于空白组，差异均有统计学意义（均P〈0．05）。与照射组和药物组比较，联合组p-EGFR和p-DNA．PKcs蛋白的表达水平均明显降低（均P〈0．05），而γH2AX蛋白表达水平均明显升高（均P〈0．05）。结论尼妥珠单抗可提高食管癌ECA-109和TE-13细�Objective To investigate the effects of nimotuzumab on radiosensitivity of ECA-109 and TE-13 esophageal carcinoma cell lines and explore its possible mechanism. Methods The ECA-109 and TE-13 cells were divided into control group, irradiation group, medicine group, and combined group （irradiation ＋ medicine）. In the combined group, ECA-109 and TE-13 cells were treated with nimotuzumab for 24 h before irradiation, and the cells were collected 2 h after irradiation. The radiosensitizing effects ofnimotuzumab on ECA-109 and TE-13 cells were evaluated by clone formation assay. Cell apoptosis was detected by flow cytometry. Western blotting was used to evaluate the expression of EGFR, p-EGFR, DNA- PKcs, p-DNA-PKcs and γH2AX. Results The values of Dq（ quasithreshold dose） , Do（ mean lethal dose） and SF2（ surviving fraction at 2 Gy） of ECA-109 and TE-13 cells in the combined group were significantly lower than those of the radiation group （ for ECA-109 cells, 1.11 vs. 1.72, 1.40 vs. 2.14, 0.42 vs. 0.66, respectively; for TE-13 cells, 0.41 vs. 0.46, 0.43 vs. 0.65, 0.40 vs. 0.71, respectively （all P〈0.05）. The sensitivity enhancement ratio （SER） of ECA-109 and TE-13 cells were 1.35 and 1.43, respectively. Flow cytometry showed that the apoptosis rate of ECA-109 and TE-13 cells in the combined group were significantly higher than those of the radiation group [ for ECA- 109 cells, （ 41.31 ± 1.52 ） % vs. （ 9.54 ±0.52） % ; for TE-13 cells, （46.28±0.28） % vs. （11.32±0.31） %, both P〈0.01 ]. Western blotting showed that the expression levels of EGFR and DNA-PKcs were not significantly different in all groups （ all P〉 0.05）. Compared with those of the control group, p-EGFR and p-DNA-PKcs of the radiation group were significantly higher in both cell lines （P〈0.05） , and the γH2AX levels in the radiation group and medicine group were significantly higher than that of the control group （P〈0.05）. Compared with those of the radiation group and medicine group, p-EGF

关 键 词：食管肿瘤 受体 表皮生长因子 辐射耐受性 尼妥珠单抗 

分 类 号：R735.1[医药卫生—肿瘤]