^(131)I-Herceptin联合高能X线对HER2高表达人乳腺癌细胞的协同杀伤及机制研究  

Synergism effect of ^(131)I-Herceptin and high-energy X-ray on HER2 overexpressed breast cancer cells

作　　者：张迎[1] 袁胜利[2] 郑勤[1] 张全安[1] 徐瀚峰[1] 

机构地区：[1]南京市第二医院肿瘤科,江苏南京210003 [2]山东省青岛市市立医院肿瘤二科,山东青岛266011

出　　处：《中国生化药物杂志》2015年第9期44-47,共4页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的研究131I-Herceptin联合高能X线对HER2高表达人乳腺癌SK-BR-3细胞的协同杀伤及机制。方法运用免疫组化法、荧光原位杂交法(fluorescence in situ hybridization,FISH)检测SK-BR-3细胞的人类表皮生长因子受体2(human epidermal growth factor receptor-2,HER2)蛋白表达和基因扩增,采用Iodogen法制备131I-Herceptin,MTT法筛选131I-Herceptin杀伤SK-BR-3细胞的IC15浓度。根据是否应用131I-Herceptin分为对照组及加药组,分别给予高能X线(0、2、4、6 Gy),以克隆形成试验分析协同杀伤效应;将细胞分为空白对照组、药物组(131I-Herceptin)、高能X线组(2Gy)、联合组(131I-Herceptin+2Gy),以A0/EB法检测细胞凋亡率及死亡率,以流式细胞仪检测各组细胞周期。结果 SK-BR-3细胞为HER2高表达细胞。131I-Herceptin的标记率为86.8%,放射化学纯度为93.9%,放射性比活度为868.3μci/mg。131I-Herceptin的IC15为15.625μci/m L。发现131I-Herceptin(F=628.008,P<0.05)及高能X线(F=964.970,P<0.05)对SK-BR-3细胞均有显著的杀伤作用,可明显降低细胞的SF值,且2者具有交互作用(F=113.046,P<0.05),即2者协同降低SF值。空白组、外照射组、药物组及联合组细胞凋亡率与死亡率比较差异均有统计学意义(F=103.324,F=13.330,均P<0.05),且两两比较差异均有统计学意义(P<0.05);经外照射及131I-Herceptin联合作用后,细胞周期均发生明显改变,由G1期向G2期和S期转移。结论131I-Herceptin联合高能X线对HER2高表达人乳腺癌SK-BR-3细胞具有协同杀伤作用。Objective To study the synergism effect of131I-Herceptin and high-energy X-ray on HER2 overexpressed breast cancer SK-BR-3 cells.Methods The protein expression and gene amplification of human epidermal growth factor receptor-2（ HER2） in SK-BR-3 cells were identified by immunohistochemistry and fluorescence in situ hybridization（ FISH） method,131I-Herceptin was prepared by iodogen method, and the IC15 concentration of131I-Herceptin on SK-BR-3 cell were selected by MTT method. The cells were divided into control group and drug group according to131 IHerceptin used or not,and were delivered five different doses of external irradiation（ 0,2,4 and 6Gy）,and the synergism effect was detected by colonogenic assay. The cells were divided into blank group,drug group（131I-Herceptin）,X-ray group（ 2 Gy external irradiation） and combination group（131I-Herceptin ＋ 2 Gy external irradiation）,the apoptosis rate and death rate were detected by AO / EB method and cell cycle were detected by flow cytometry. Results The labling rate,radiochemical purity and specific radioactivity of131I-Herceptin were 86. 8%,93. 9% and 868. 3 μci / mg,respectively. The IC15 of131I-Herceptin was 15. 625 μci / m L.131I-Herceptin and high-energy X-ray significantly reduced surviving fraction（ SF）（ F =628. 888,F = 964. 97,P〈0. 05） and there were interactions between them（ F = 113. 046,P〈0. 05）. There were significant differences in apoptosis rate and death rate among blank group,drug group,X-ray group and combination group（ F = 103. 324,F = 13. 33,all P〈0. 05）,and there were significant differences of pairwise comparison（ P〈0. 05）. After irradiation and131I-Herceptin administration,the cell cycle changed obviously from G1-phase to G2- and S-phase. Conclusion131I-Herceptin combined with high-energy X-ray has the synergism effect on HER2 overexpressed breast cancer SK-BR-3cells.

关 键 词：乳腺癌 人表皮生长因子受体2 放射免疫治疗 协同杀伤 

分 类 号：R737.9[医药卫生—肿瘤]