达沙替尼对曲妥珠单抗杀伤胃癌细胞BGC-823增敏作用的体外实验

Dasatinib enhances the sensitivity of gastric cancer cell BGC-823 to trastuzumab

目的检测蛋白激酶c-Src抑制剂达沙替尼(Dasatinib)对曲妥珠单抗(Trastuzumab)杀伤胃癌细胞的增敏作用。方法从胃癌细胞系BGC-823、SGC-7901、MGC-803以及NCI-N87中筛选HER2与c-Src高表达的细胞系。最终筛选得到BGC-823细胞,使用系列浓度梯度的达沙替尼和曲妥珠单抗孵育BGC-823细胞48 h,利用MTT法进行药物作用的剂量-效应关系实验,计算药物杀伤细胞的抑制率,最终确定并选取达沙替尼和曲妥珠单抗作用于BGC-823的浓度。在此基础上,利用细胞增殖实验与凋亡实验检测达沙替尼是否能够上调曲妥珠单抗对BGC-823细胞的杀伤作用。结果达沙替尼杀伤BGC-823的起效作用浓度(IC25)为(0.47±0.10)μmol/L;曲妥珠单抗杀伤BGC-823细胞的半数作用浓度(IC50)为(2.25±0.33)μg/ml。达沙替尼能够上调曲妥珠单抗对BGC823细胞的杀伤作用:使用达沙替尼预处理BGC-823细胞能够降低曲妥珠单抗作用的IC50值[从(2.30±0.22)μg/ml下调为(0.31±0.05)μg/ml]。结论达沙替尼能够上调曲妥珠单抗对胃癌细胞的杀伤作用。

Objective To reveal whether dasatinib, an inhibitor of kinase c-Src, enhances the sensitivity of BGC-823 cells to trastuzumab. Methods The level of endogenous proteins c-Src and HER2 in gastric cancer cell lines BGC-823, SGC-7901, MGC-803, and NCI-N87 were detected by western blot. The concentration-effect curves of dasatinib and trastuzumab were determined by MTT assay. The regulatory role of dasatinib on trastuzumab to BGC-823 cells was identified by MTT, colony formation and apoptosis assays. Results Dasatinib and trastuzumab inhibited BGC-823 cells survival in a dose-dependent manner. The IC25（25% inhibitory effect concentration） of dasatinib was 0.47±0.10μmol/L and IC50（50% inhibitory effect concentration） of trastuzumab was 2.25±0.33μg/ml. Treatment of 0.2μmol/L dasatinib enhanced the effect of trastuzumab on BGC-823, and the IC50 of trastuzumab decreased from 2.30±0.22μg/ml to 0.31±0.05μg/ml. Conclusion Dasatinib enhances the effect of trastuzumab on BGC-823 cells.