CL-387785对非小细胞肺癌H1975细胞侵袭转移的抑制及放疗增敏作用研究  

The inhibitory effect of CL-387785 on the invasion and metastasis of T790M EGFR mutant H1975 cells and its sensitization of radiotherapy

作　　者：盛兆瑛[1] 蔡勇[1] 倪健[2] 

机构地区：[1]同济大学附属上海市肺科医院放疗科,上海200433 [2]上海市肺科医院肿瘤科,200433

出　　处：《临床肿瘤学杂志》2015年第7期577-582,共6页Chinese Clinical Oncology

摘　　要：目的探讨不可逆EGFR抑制剂CL-387785对EGFR T790M突变非小细胞肺癌H1975细胞侵袭转移能力的抑制及放疗增敏作用。方法分别采用0、10、25、50、100 nmol/L CL-387785处理H1975细胞24、48、72和96 h后用四甲基偶氮唑盐（MTT）比色法检测增殖抑制率变化,同时用流式细胞术Annexin-FITC/PI双染法检测不同浓度CL-387785处理48、96 h后的细胞凋亡情况,Transwell法及划痕实验检测不同浓度CL-387785处理48、96 h后的H1975细胞侵袭及迁移能力变化,采用克隆形成实验检测CL-387785处理的H1975细胞经不同剂量X线（0、2、4、6、8和10 Gy）照射的存活分数（SF）并通过单击多靶模型拟合细胞存活曲线计算增敏比（SER）。结果 CL-387785可抑制H1975细胞增殖,且在10~100 nmol/L范围内随作用时间延长,增殖抑制率升高,总体上抑制效应呈浓度和时间依赖方式;CL-387785可呈浓度依赖的方式诱导H1975细胞凋亡并缩短细胞迁移距离和减少穿膜细胞数（P〈0.05）;在2~10 Gy照射范围内,10、25、50、100 nmol/L CL-387785处理后的SF均低于0 nmol/L,且SF随CL-387785浓度的增加而降低,以上差异均有统计学意义（P〈0.05）;相对于0 nmol/L,10、25、50、100 nmol/L CL-387785处理后的SER依次为1.17、1.39、2.88和3.64。结论不可逆EGFR抑制剂CL-387785可抑制EGFR T790M突变非小细胞肺癌H1975细胞的侵袭转移能力并具有放射增敏作用。Objective To investigate the inhibitory effect of CL-387785 on the invasion and metastasis of EGFR T790M mu- tant H1975 cells and its sensitization of radiotherapy. Methods The H1975 cell was treated with 0, 10, 25, 50, 100 nmol/L CL- 387785. The MTT colorimetric assay was used to detect the proliferation inhibition rates at 24, 48, 72 and 96 h after treatment. Moreo- ver, Annexin-FITC/PI double staining method via flow cytometry was used to detect the apoptotic rates of different concentrations of CL-387785 at 48 and 96 h. Changes of invasion and migration of H1975 cells treated with CL-387785 were detected by Transwell and scratch assay, respectively. The clone formation assay was employed to investigate the survival fraction （SF） of the H1975 ceUs after exposure to CL-387785 following different doses of X-ray （0, 2, 4, 6, 8 and 10 Gy） irradiation. The target model was fitted to the cell survival curve to calculate the sensitization enhancement ratio （SER）. Results CL-387785 inhibited the proliferation of H1975 cells, and dose- and time-dependently increased the inhibition rates of the proliferation in range of 10-100 nmol/L. CL-387785 could induce the apoptosis and decrease the number of penetrate-membrane cell and migration distance of H1975 ceils in a dose-dependent manner （P〈0. 05）. In the 2-10 Gy irradiation range, the SF of 10, 25, 50 and 100 nmol/L CL-387785 were lower than that of 0 nmol/L, and SF decreased with the increase of concentration of CL-387785 and above differences were statistically significant （P〈0. 05）. Compared with the 0 nmol/L CL-387785, the SER of 25, 50, 10, 100 nmol/L CL-387785 SER after treatment were 1.17, 1.39, 2. 88 and 3.64, respectively. Conclusion The irreversible EGFR inhibitor CL-387785 can inhibit the invasion and metastasis of T790M EGFRmutant H1975 cells and has the effect of radiation sensitization.

关 键 词：EGFR抑制剂 非小细胞肺癌 侵袭转移 放疗增敏 

分 类 号：R734.2[医药卫生—肿瘤]