PTEN低表达增加胃癌曲妥珠单抗的耐药性  

The low expression of PTEN increased resistance to trastuzumab in gastric cancer

作　　者：吕志诚[1] 党政[1] 任彦顺[1] 王湘辉[1] 

机构地区：[1]兰州军区兰州总医院肝胆外科,甘肃兰州730050

出　　处：《现代肿瘤医学》2016年第16期2512-2516,共5页Journal of Modern Oncology

基　　金：甘肃省科技支撑计划项目(编号:1104FKCA154)

摘　　要：目的：明确PTEN活性和胃癌曲妥珠单抗耐药性的关系。方法：用Western blot方法检测胃腺癌细胞株中HER2蛋白、PTEN蛋白、PI3K蛋白、AKT蛋白的表达;用CCK-8方法检测曲妥珠单抗对HER2阳性/PTEN野生型NCI-N87细胞的生长抑制作用;构建PTEN siRNA载体并将其PTEN siRNA载体感染HER2阳性/PTEN野生型NCI-N87细胞,建立稳定表达PTEN siRNA的细胞株N87-PD和阴性对照N87-NC,用Western blot验证其表达水平;同时用CCK-8验证曲妥珠单抗对各组细胞生长的影响。结果：Western blot结果显示,NCI-N87细胞的HER2表达水平高于MNK-7细胞。CCK-8检测表明,随着药物浓度的增加,细胞存活率持续呈下降趋势。三种siRNA中以siRNA-1对PTEN mRNA的干扰效果最优,选择siRNA-1进行后续实验。实时定量PCR法测定最终病毒滴度为1.56×10~9/ml（siRNA-1）和1.62×10~9/ml（NC）。PTEN siRNA对NCI-N87细胞的PTEN蛋白表达有明显的下调作用且N87-PD细胞中磷酸化AKT蛋白水平显著高于对照组细胞N87-NC;曲妥珠单抗作用72h后,CCK-8测定N87-NC细胞存活率显著下降,而N87-PD细胞存活率与阴性对照组细胞无显著差异。结论：PTEN低表达增加胃癌曲妥珠单抗的耐药性。Objective：To study the relationship between PTEN activity and trastuzumab resistance of gastric canc-er. Methods：To use Western blotting method to detect the expression of HER2 protein in gastric cancer cell lines, PTEN protein,PI3K protein,AKT protein. CCK - 8 was used to detect the growth of trastuzumab for HER2 positive/PTEN wild - type NCI - N87 cells. To construct PTEN siRNA lentiviral vector and infect it to the NCI - N87 cell which carry HER2 positive/ PTEN protein wild - type for establishing stable expression PTEN siRNA control cell lines N87 - PD and negative lines N87 - NC. Western blotting was used to detect the inhibition efficiency and CCK - 8 de-tect cell survival rate after trastuzumab treatment. Results：Western blot results showed that the expression level of NCI - N87 in HER2 cells was higher than that in MNK - 7 cells. CCK - 8 test showed that the survival rate of the cells was decreased with the increase of drug concentration. siRNA - 1 PTEN mRNA interference effect was optimal in the three kinds of siRNAs,and siRNA - 1 was selected for the following experiments. The final viral titer was 1. 56 ＆#215; 109 / ml（siRNA - 1）and 1. 62 ＆#215; 109 / ml（NC）by Real time quantitative PCR assay. PTEN - siRNA on NCI - N87 cells PTEN protein expression was significantly down regulated and phosphorylated AKT protein levels in N87 - PD cells was significantly higher than that of the control group cells N87 - NC. CCK - 8 assay showed that N87 - NC cells survival rate decreased significantly and N87 - PD cell survival rate and negative cells in the control group no signifi-cant difference after trastuzumab treatment for 72h. Conclusion：The low expression of PTEN increased resistance to trastuzumab in gastric cancer.

关 键 词：PTEN 胃癌 曲妥珠单抗 活性 耐药性 

分 类 号：R735.2[医药卫生—肿瘤]