CAJ | 학술논문

目的：研究131 I-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131 I-Herceptin，MTT法筛选131 I-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。131 I-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／mL。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131 I-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。
목적：연구131 I-Herceptin연합고능X선대HER2고표체인유선암SK-BR-3세포적협동살상급궤제。방법운용면역조화법、형광원위잡교법（fluorescence in situ hybridization，FISH）검측SK-BR-3세포적인류표피생장인자수체2（human epidermal growth factor receptor-2，HER2）단백표체화기인확증，채용Iodogen법제비131 I-Herceptin，MTT법사선131 I-Herceptin살상SK-BR-3세포적IC15농도。근거시부응용131I-Herceptin분위대조조급가약조，분별급여고능X선（0、2、4、6 Gy），이극륭형성시험분석협동살상효응；장세포분위공백대조조、약물조（131I-Herceptin）、고능X선조（2Gy）、연합조（131I-Herceptin＋2Gy），이A0／EB법검측세포조망솔급사망솔，이류식세포의검측각조세포주기。결과 SK-BR-3세포위HER2고표체세포。131 I-Herceptin적표기솔위86.8％，방사화학순도위93.9％，방사성비활도위868.3μci／mg。131I-Herceptin적IC15위15.625μci／mL。발현131I-Herceptin（F＝628.008，P＜0.05）급고능X선（F＝964.970，P＜0.05）대SK-BR-3세포균유현저적살상작용，가명현강저세포적SF치，차2자구유교호작용（F＝113.046，P＜0.05），즉2자협동강저SF치。공백조、외조사조、약물조급연합조세포조망솔여사망솔비교차이균유통계학의의（F＝103.324，F＝13.330，균P＜0.05），차량량비교차이균유통계학의의（P＜0.05）；경외조사급131I-Herceptin연합작용후，세포주기균발생명현개변，유G1기향G2기화S기전이。결론131 I-Herceptin 연합고능X선대HER2고표체인유선암SK-BR-3세포구유협동살상작용。
Objective To study the synergism effect of 131 I-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, 131 I-Herceptin was prepared by iodogen method, and the IC15 concentration of 131 I-Herceptin on SK-BR-3 cell were selected by MTT method.The cells were divided into control group and drug group according to 131 I-Herceptin 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 of 131 I-Herceptin were 86.8%, 93.9% and 868.3 μci/mg, respectively.The IC15 of 131 I-Herceptin was 15.625μci/mL.131 I-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 and 131I-Herceptin administration, the cell cycle changed obviously from G1-phase to G2-and S-phase.Conclusion 131 I-Herceptin combined with high-energy X-ray has the synergism effect on HER2 overexpressed breast cancer SK-BR-3 cells.