131I-Trastuzumab对人表皮生长因子受体2过表达乳腺癌细胞的杀伤效应及机制研究  

Tumor inhibitory effects of 131I-Trastuzumab on human epidermal growth factor receptor 2 overex- pressing breast cancer cells and its possible mechanisms

作　　者：张龙杰[1] 侯和磊[1] 王国明[2] 韩真真[1] 张晓春[1] 袁胜利[1] 

机构地区：[1]青岛大学附属市立医院肿瘤科,266000 [2]青岛大学附属医院核医学科

出　　处：《中华核医学与分子影像杂志》2015年第4期293-297,共5页Chinese Journal of Nuclear Medicine and Molecular Imaging

摘　　要：目的观察131I-Trastuzumab对人表皮生长因子受体2（HER2）过表达乳腺癌细胞的杀伤效应并探讨其机制。方法（1）免疫荧光法检测乳腺癌细胞（BT474、MCF-7和HCC1937）表面HER2表达水平。（2）Iodogen法和超滤法制备并纯化131I-Trastuzumab，测定其标记率、放化纯及免疫结合率。（3）细胞计数试剂盒-8（CCK-8）法检测不同剂量131I、Trastuzumab和131I-Trastuzumab对HER2过表达乳腺癌BT474细胞的杀伤效应并进行浓度筛选。（4）Westernblot检测对照组及各干预组中总Akt及磷酸化Akt（p-Akt）表达水平。统计学分析采用单因素方差分析、析因设计资料的方差分析、Bonferroni校正法及Pearson相关分析。结果（1）乳腺癌BT474细胞HER2表达水平明显高于MCF-7和HCC1937细胞。（2）131I-Trastuzumab的标记率、放化纯和免疫结合率分别为（89．71±2．93）％、（91．80±1．43）％和（58．84±3．35）％。（3）131I、Trastuzumab及131I-Trastuzumab对BT474细胞的杀伤效应具有剂量依赖性（r=-0．964．-0．912和-0．618，均P〈0．05）；对照组、131I（4．625GBq／L）、Trastuzumab（125．0ms／L）及131I-Trastuzumab（4．625GBq／L）干预后的BT474细胞存活率分别为（100．00±4．54）％、（64．36±1．51）％、（58．09±4．14）％和（34．73±5．03）％，131I-Trastuzumab干预后的细胞存活率明显低于相应的”。I和Trastuzumab干预组（t=10．373和8．180，均P〈0．05）。131I、Trastuzumab均可杀伤BT474细胞（F=213．326和313．564，均P〈0．05），两者间存在协同作用（F=9．226，P〈0．05；CDI=0．929）。（4）Westernblot结果显示对照组、131I、Trastuzumab和131I-Trasmzumab组之间总Akt的表达量差异无统计学意义（F=0．208，P〉O．05）；与对照组及131I组相比，Trastuzumab、131I-Trastuzumab组p-Akt含量明显降低（t=12．524、15．984、7．347和10．807，均P〈0．05），而Trastuzumab与131I-Trastu—zumab组之Objective To explore antitumor effect of 131I-Trastuzumab on human epidermal growth factor receptor（HER） 2 overexpressing breast cancer ceils and investigate its possible mechanism. Methods The expression levels of HER2 detected with of three different breast cancer cell ：e. Trastuzumab was labeled with 131 lines （BT474, MCF-7, HCC1937） were I using the Iodogen method and 1311-Tras- tuzumab was isolated with uhrafiltration membrane, then the labeling efficiency, radiochemical purity and immunoreactivity were measured. The effects of 131I, Trastuzumab and 131I-Trastuzumab on viability of BT474 cells were evaluated with cell counting kit-8 （CCK-8） assay. The levels of total Akt and phosphoryla- ted Akt （p-Akt） were detected with Western blot analysis. One-way analysis of variance （ANOVA）, ANOVA for factorial design, Bonferroni correction and Pearson correlation analysis were used for data analysis. Resuits The expression level of HER2 in BT474 ceils was much higher than those in HCC1937 and MCF-7 cells. The labeling efficiency, radiochemical purity and immunoreactivity of 131I-Trastuzumab were （89.71± 2.93）%, （91.80±1.43）% and （58.84±3.35）% respectively. 131I （4.625 GBq/L）, Trastuzumab（ 125.0mg/L） and 131I-Trastuzumab（ 4.625 GBq/L） exhibited a dose-dependent cytotoxicity against BT474 cells （r=-0.964, -0.912, -0.618 ; all P〈0.05）. The cell viability of 131i.Trastuzumab treated gourp （ 34.73% ±5.03% ） was significantly lower than those of 131I and Trastuzumab treated groups （64.36% ± 1.51% and 58.09% ±4.14% ; t= 10.373 and 8.180, both P〈0. 05）, and the cell viability of control group was （100.00±4.54）%. 131I- Trastuzumab shown a positive multiplicative interaction between 131I and Trastuzumab （F= 9.226, P〈0.05; CDI = 0.929）. Western blot results showed that there was no significant difference of total Akt expression a- mong the control group, 131I group, Trastuzumab group and 131I-Trastuzmab group （ F = 0. 208, P〉0.05 ）. P-Akt expre

关 键 词：乳腺肿瘤 受体 表皮生长因子一尿抑胃素 抗体 单克隆 碘放射性同位素 肿瘤细胞 培养的 

分 类 号：R737.9[医药卫生—肿瘤]