摘要
目的 研究野黄芩苷对胃癌HGC-27细胞增殖、凋亡、迁移、侵袭及铁死亡的影响,探讨铁死亡调控机制.方法 CCK8检测细胞活力;Hochest检测细胞凋亡;细胞划痕和Transwell检测细胞迁移和侵袭;DCFH-DA检测细胞ROS;JC-1检测细胞线粒体膜电位;Ferro Orange检测细胞Fe2+.在线数据库获取野黄芩苷、胃癌和铁死亡靶点,并对交集靶点进行蛋白互作分析.Western Blotting检测野黄芩苷干预细胞后的EGFR和COX2表达.结果 野黄芩苷对HGC-27细胞24 h和48 h的IC50分别为87.08和61.69µM.野黄芩苷能够明显诱导细胞凋亡、ROS、线粒体膜电位极化、Fe2+和COX2 表达水平(P＜0.05),明显抑制细胞划痕愈合、迁移、侵袭和EGFR表达水平.野黄芩苷、胃癌和铁死亡的交集靶点为EGFR、COX2、MAPK14、HRAS、NOX4、CA9、ALOX5 和 KDM5A.野黄芩苷能够有效逆转 NSC 228155 和 Rofecoxib 对HGC-27细胞活力及Fe2+水平的影响(P＜0.05).结论 野黄芩苷能够抑制胃癌HGC-27细胞活力、迁移和侵袭,以及诱导其凋亡和铁死亡.其中,野黄芩苷诱导铁死亡的机制与其调控EGFR/COX2信号通路有关.
Abstract
Objective To explore the impact of scutellarin on proliferation,apoptosis,migration,invasion,and ferroptosis processes of gastric cancer HGC-27 cells.Methods Scutellarin was used to intervene with HGC-27 cells.Cell viability was assessed using CCK8,apoptosis was detected by Hochest staining,migration and invasion using scratch assays and Transwell,ROS were measured using DCFH-DA,mitochondrial membrane potential was assessed using JC-1 staining,and Fe2+levels were tested by Ferro Orange.Scutellarin,gastric cancer,and ferroptosis targets were obtained from online databases,and protein-protein interaction analysis was conducted on intersecting targets.Western Blotting was performed to evaluate the expression of EGFR and COX2 after scutellarin intervention.Results The IC50 of scutellarin on HGC-27 cells at 24 h and 48 h were 87.08 pM and 61.69 pM,respectively.Scutellarin significantly induced apoptosis,ROS,mitochondrial membrane potential polarization,Fe2+and COX2 expression levels,while inhibiting scratch healing,migration,invasion,and EGFR expression compared to blank HGC-27 cells,all with statistical significance(P＜0.05).Intersecting targets of scutellarin,gastric cancer,and ferroptosis included EGFR,COX2,MAPK14,HRAS,NOX4,CA9,ALOX5 and KDM5A.Scutellarin effectively reversed the impact of NSC 228155 and rofecoxib on HGC-27 cell viability and Fe2+levels,both statistically significant(P＜0.05).Conclusion Scutellarin demonstrated the ability to inhibit the viability,migration,and invasion of gastric cancer HGC-27 cells,while also inducing apoptosis and ferroptosis in these cells.Notably,scutellarin-induced ferroptosis mechanism is related to its modulation of the EGFR/COX2 signaling pathway.
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