抑制SHP2和FGFR2调控RAS/ERK及PI3K/AKT通路治疗FGFR2融合胃癌
Co-inhibiting SHP2 and FGFR2 to treat FGFR2-fused gastric cancer by regulating RAS/ERK and PI3K/AKT signaling pathways
张玥 1汪越 1魏禹焘 2禹立霞 1刘宝瑞 1魏嘉1
作者信息
- 1. 南京大学医学院附属鼓楼医院肿瘤科(南京市 210008)
- 2. 南京中医药大学中西医结合鼓楼临床医学院
- 折叠
摘要
目的:探究共抑制成纤维细胞生长因子受体2(fibroblast growth factor receptor 2,FGFR2)和Src同源2结构域的蛋白酪氨酸磷酸酶 2(Src homology region 2-containing protein tyrosine phosphatase 2,SHP2)在FGFR2融合胃癌中的应用前景与作用机制.方法:构建过表达TACC2-FGFR2融合基因与对照慢病毒载体的人胃癌细胞系MKN45TACC2-FGFR2、MKN45NC、NUGC4TACC2-FGFR2、NUGC4NC,分别用FGFR2抑制剂AZD4547、SHP2抑制剂SHP099或联药进行处理,通过细胞计数试剂盒(CCK-8)、划痕实验检测肿瘤细胞的增殖、迁移能力.以不同处理方式作用于MKN45TACC2-FGFR2、MKN45NC1 h或 48 h后,采用Western blot法检测FGFR2、SHP2以及下游RAS/ERK、PI3K/AKT信号通路变化.结果:在MKN45TACC2-FGFR2 与NUGC4TACC2-FGFR2 中联用AZD4547与SHP099可以比单药更显著地抑制肿瘤细胞的增殖与迁移.药物处理 1 h后,相较于AZD4547单药,联药在MKN45TACC2-FGFR2中进一步抑制了RAS/ERK、PI3K/AKT信号通路.药物处理 48 h与 1 h相比,AZD4547单药组中磷酸化FGFR与磷酸化SHP2出现了反馈性激活,且始终不能抑制RAS/ERK通路,但联药组可以持续地抑制上游的FGFR2、SHP2信号以及下游的RAS/ERK、PI3K/AKT通路.结论:共抑制FGFR2和SHP2可以通过下调RAS/ERK及PI3K/AKT通路有效抑制FGFR2融合胃癌,为FG-FR2融合突变胃癌患者带来新的治疗模式.
Abstract
Objective:In this study,we explored the application prospects and mechanisms of action of co-inhibiting fibroblast growth factor receptor 2(FGFR2)and Src homology region 2-containing protein tyrosine phosphatase 2(SHP2)in gastric cancer with the TACC2-FGFR2 fu-sion gene.Methods:We established human gastric cancer cell lines overexpressing the TACC2-FGFR2 fusion gene(MKN45TACC2-FGFR2 and NUGC4TACC2-FGFR2 cells)or a control lentiviral virus(MKN45NC and NUGC4NC cells).The cells were treated with the FGFR2 inhibitor AZD4547,the SHP2 inhibitor SHP099,or a combination of both.The proliferation and migration of tumor cells were detected using cell counting Kit-8(CCK-8)and scratch assays.After treating MKN45TACC2-FGFR2 and NUGC4TACC2-FGFR2 cells with different formulations for 1 or 48 h,Western blot was used to detect variations in the levels of FGFR2,SHP2,and proteins downstream of the RAS/ERK and PI3K/AKT signaling pathways.Results:Com-pared to monotherapy,the combination of AZD4547 and SHP099 significantly inhibited the proliferation and migration of MKN45TACC2-FGFFR2 and NUGC4TACC2-FGFFR2 cells.After 1 h of treatment,the combination therapy inhibited the RAS/ERK and PI3K/AKT pathways in MKN45TACC2-FGFFR2 cells to a greater extent than the AZD4547 monotherapy.Forty-eight hours of AZD4547 monotherapy resulted in feedback activation of p-FGFR and p-SHP2,but failed to inhibit the RAS/ERK pathway.However,combination therapy continuously suppressed upstream FGFR2 and SHP2 signaling,as well as downstream RAS/ERK and PI3K/AKT pathways.Conclusions:Co-inhibiting FGFR2 and SHP2 further inhibit gastric cancer with the TACC2-FGFR2 fusion gene by suppressing the RAS/ERK and PI3K/AKT pathways.These findings provide a new treatment mode for patients with gastric cancer with the TACC2-FGFR2 fusion gene.
关键词
胃癌/靶向治疗/融合基因/纤维细胞生长因子受体2/Src同源2结构域的蛋白酪氨酸磷酸酶2Key words
gastric cancer/targeted therapy/gene fusion/fibroblast growth factor receptor 2(FGFR2)/Src homology region 2-contain-ing protein tyrosine phosphatase 2(SHP2)引用本文复制引用
基金项目
国家自然科学基金(82073382)
江苏省基础研究计划自然科学基金(BK20230151)
出版年
2024
NETL
NSTL
万方数据