摘要
目的探索外源性人表皮生长因子受体2(HER2)基因导入使HER2阴性胃癌细胞系对曲妥珠单抗敏感性的影响。方法构建人HER2真核表达载体,转染人HER2阴性胃癌细胞系(HGC-27和MGC803)及正常胃上皮细胞GES1(作为对照),细胞免疫荧光与蛋白印迹(Western blot,WB)检测HER2表达及下游信号通路变化,CCK-8法检测细胞增殖活性。结果 HER2阴性胃癌细胞及正常细胞导入外源性HER2后,HER2蛋白表达明显上调,但磷酸化HER2(pHER2)表达无变化。外源性HER2基因导入后,HER2阴性细胞对曲妥珠单抗的敏感性无增加,甚至有下降趋势。此外,外源性HER2基因导入后,HER2下游PI3K/AKT、MAPK通路不但未激活,反而受到明显抑制。结论外源性HER2基因导入无法使HER2阴性胃癌从曲妥珠单抗治疗中获益,主要原因可能是外源性HER2基因无法使其下游PI3K/AKT、MAPK通路激活,外源性和内源性HER2发挥的作用可能不同,有待后续深入研究。
Objective To explore the feasibility of HER2-negative gastric cancer cell lines benefiting from trastuzumab treatment after exogenous HER2 gene introduction. Methods Human HER2 eukaryotic expression vector was constructed and transfected into human gastric cancer cell lines(HGC-27 and MGC803) and normal gastric epithelial cell GES1(as control). Immunofluorescence and Western blot were used to detect HER2 and its downstream signaling pathways, and CCK-8 assay was used to measure the viability of cells. Results After the introduction of exogenous HER2 into HER2-negative gastric cancer cells and normal cells, HER2 was significantly upregulated rather than phosphorylated HER2(pHER2). Also, the sensitivity of HER2-negative cells to trastuzumab wasn’t increased after the introduction of exogenous HER2. In addition, PI3 K/AKT and MAPK pathways were not activated but significantly inhibited after the introduction of exogenous HER2. Conclusions Exogenous HER2 introduction cannot benefit HER2-negative gastric cancer from trastuzumab treatment. The main reason might be that exogenous HER2 does not activate its downstream PI3 K/AKT and MAPK pathways. The role of exogenous and endogenous HER2 may be different,which will be explored in further study.
作者
张朦琦
陈祖华
杨菁
章程
沈琳
高静
ZHANG Meng-qi;CHEN Zu-hua;YANG Jing;ZHANG Cheng;SHEN Lin;GAO Jing(Key Laboratory of Carcinogenesis and Translational Research,Ministry of Education,Department of Gastrointestinal Oncology,Peking University Cancer Hospital&Institute,Beijing 100142,China)
出处
《基础医学与临床》
CSCD
2019年第4期489-494,共6页
Basic and Clinical Medicine
基金
国家重点研发计划(2017YFC1308900)
