摘要
目的探讨葛根素对胶质瘤细胞增殖、迁移和凋亡的影响及机制。方法GEO数据库分析胶质瘤与线粒体疾病共相关差异性表达基因;CCK-8检测细胞活力;划痕和Transwell检测细胞迁移;EdU检测细胞增殖;TUNEL检测细胞凋亡;Mito-Tracker检测线粒体膜电位;ATP试剂盒分析ATP含量;免疫印迹分析蛋白质表达;皮下异种移植肿瘤模型分析葛根素抗肿瘤效果。结果胶质瘤和线粒体疾病共相关差异性表达基因178个;经GO和KEGG富集及相互作用网络分析,筛选出12个核心基因;其中泛素蛋白C(ubiquitin C,UBC)在胶质瘤患者组织中高表达。葛根素可与UBC结合,并在动物和细胞水平降低UBC表达。葛根素处理可缩小胶质瘤体积,降低胶质瘤细胞增殖和迁移,促进胶质瘤细胞凋亡;同时降低线粒体膜电位和ATP含量,并下调UBC通路蛋白表达水平。结论葛根素抑制胶质瘤细胞增殖和迁移并促进其凋亡,其机制与诱发线粒体功能紊乱有关。
Aim To investigate the effects of puerarin on the proliferation,migration,and apoptosis of glioblastoma cells and the underlying mechanisms.Methods Differentially expressed genes associated with glioma and mitochondrial disease were analyzed using the GEO database.Cytotoxicity was detected by CCK-8 assay.Cell migration was detected by the scratch wound healing assay and Transwell assay.Cell proliferation was assessed by EdU assay.Apoptosis level was measured by TUNEL assay.Mitochondrial membrane potential was detected by Mito-Tracker assay.ATP contents were detected using the ATP kit.The protein expression levels were detected by Western blot.Antitumor efficacy of puerarin was analyzed using subcutaneous xenograft.Results There were 178 genes co-related differentially expressed genes in glioma and mitochondrial disease.Core genes of co-related differentially expressed genes were screened by GO and KEGG enrichment analyses,and the interaction networks.Among them,ubiquitin C(UBC)level was highly expressed in tissues of glioma patients.Puerarin could bind to UBC and reduce UBC expression at the animal and cell levels.Puerarin treatment inhibited the growth of glioma and decreased cell proliferation,migration and promoted cell apoptosis signals.Meantime,puerarin treatment also reduced mitochondrial membrane potentials and ATP contents,and down-regulated the levels of UBC related proteins.Conclusion Puerarin inhibits the proliferation,migration and promotes apoptosis of glioma cells.The mechanism of induction of mitochondrial dysfunction is involved.
作者
盛格格
冯子瀚
程聿德
朱海丽
陈绍恢
SHENG Ge-ge;FENG Zi-han;CHENG Yu-de;ZHU Hai-li;CHEN Shao-hui(School of Pharmacy,Hubei University of Science and Technology,Xianning Hubei 437100,China;School of Clinical Medicine,Xianning Medical College,Hubei University of Science and Technology,Xianning Hubei 437100,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2024年第7期1317-1324,共8页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81971066)。
