METHODS: The research utilized subcutaneous tumor models in nude mice to assess gastric cancer progression. Protein expression was detected using Western blotting, while Q-PCR examined the expression levels of lncRNA SNHG18 and miR-211-5p in MGC-803 cells. The relationship between miR-211-5p and lncRNA SNHG18 can be analyzed by dual luciferase reporter genes. The migratory ability of MGC-803 cells was determined through wound healing and transwell experiments, and cell proliferation was evaluated using a CCK-8 assay.
RESULTS: SHP2 was found to inhibit the cytotoxic effects of CAR-T cells on MGC-803 cells, and it suppressed the expression of proteins related to the ROS/JNK/NFAT4 signaling pathway in MGC-803 cells and the miR-211-5p/BRD4 axis in CAR-T cells. In addition, the proliferation, invasion and migration of MGC-803 cells were promoted, and the expression of miR-211-5p could be inhibited specifically by ncRNA SNHG18, as shown below:SHP2 in gastric cancer cells mediates the ROS/JNK/NFAT4 signaling pathway and induces lncRNA SNHG18, which, through the miR-211-5p/BRD4 axis in CAR-T cells, promotes gastric cancer growth and metastasis.
方法:本研究利用裸鼠皮下肿瘤模型评估胃癌进展。蛋白质表达检测使用蛋白质印迹,而Q-PCR检测了MGC-803细胞中lncRNASNHG18和miR-211-5p的表达水平。miR-211-5p和lncRNASNHG18之间的关系可以通过双荧光素酶报告基因分析。通过伤口愈合和transwell实验确定MGC-803细胞的迁移能力,使用CCK-8测定评估细胞增殖。
结果:SHP2被发现抑制CAR-T细胞对MGC-803细胞的细胞毒作用,抑制MGC-803细胞中ROS/JNK/NFAT4信号通路相关蛋白的表达和CAR-T细胞中miR-211-5p/BRD4轴的表达。此外,扩散,促进MGC-803细胞的侵袭和迁移,miR-211-5p的表达可以被ncRNASNHG18特异性抑制,如下所示:SHP2在胃癌细胞中介导ROS/JNK/NFAT4信号通路,通过CAR-T细胞中的miR-211-5p/BRD4轴,促进胃癌的生长和转移。