Itraconazole通过靶向C1GALT1抑制胃癌细胞侵袭和转移  

Itraconazole Inhibits Gastric Cancer Cell Invasion and Metastasis by Targeting C1GALT1

作　　者：董晓霞[1] 刘玉凤 俞靖蕾 张佳丽 孙碧瑶 李知远 沈力[1] DONG Xiao-xia;LIU Yu-feng;YU Jing-lei;ZHANG Jia-li;Sun Bi-yao;LI Zhi-yuan;SHEN Li(Institute of Basic Medical Sciences,Hubei University of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院基础医学研究所,湖北十堰442000

出　　处：《湖北医药学院学报》2022年第5期441-446,F0002,F0003,共8页Journal of Hubei University of Medicine

基　　金：国家自然科学基金项目(81902494)。

摘　　要：目的:探讨伊曲康唑(Itraconazole)对胃癌细胞侵袭转移的影响及机制。方法:体外培养胃癌AGS和MKN45细胞,Itraconazole处理后采用CCK-8和克隆形成测定细胞增殖,Transwell和划痕实验观察细胞侵袭迁移,流式细胞术检测细胞周期和凋亡。随后,利用TCGA数据库挖掘C1GALT1在胃癌组织中的表达谱,采用GSEA软件对C1GALT1进行功能注释,同时通过Western blot和Lectin blot分析Itraconazole对C1GALT1及其相关蛋白表达的影响。将C1GALT1过表达质粒转染至AGS和MKN45细胞,Transwell小室检测Itraconazole作用后细胞侵袭和迁移能力的变化。结果:功能实验发现,Itraconazole抑制胃癌细胞的增殖、侵袭和迁移,诱导G0/G1期阻滞,促进凋亡。生物信息学分析显示,C1GALT1在胃癌组织中高表达,且与O-糖基化途径及自噬通路密切相关。机理研究表明,Itraconazole下调胃癌细胞中C1GALT1的蛋白表达,并可阻断O-糖基化,提高自噬水平。过表达C1GALT1,Itraconazole对胃癌细胞侵袭和迁移的抑制作用减弱。结论:Itraconazole可通过直接靶向调节C1GALT1抑制胃癌细胞的侵袭和转移,在胃癌中具有抗肿瘤活性。Objective To explore the effects and mechanisms of itraconazole on the invasion and metastasis of gastric cancer cells.Methods Human gastric cancer cell lines AGS and MKN45 were cultured in vitro.After treatment with itraconazole,CCK-8 and colony formation assays were used to measure cell proliferation.Transwell and scratch assays were utilized to observe cell invasion and migration.Flow cytometry was conducted to detect the cell cycle and apoptosis.Subsequently,C1 GALT1 expression profile in gastric cancer tissues was investigated The Cancer Genomes Atlas(TCGA)database,and the biological function of C1 GALT1 was annotated by Gene Set Enrichment Analysis(GSEA).Western blot and Lectin blot assays were applied to determine whether itraconazole could influence the expression of C1 GALT1 and its related proteins.After transfection with the C1 GALT1 overexpression plasmid,the changes in invasive and migratory abilities of AGS and MKN45 cells were examined by transwell assay following itraconazole treatment.Results Functional experiments showed that itraconazole suppressed cell proliferation,invasion and migration,induced cell cycle arrest at G0/G1 phase,and promoted cell apoptosis in gastric cancer.Bioinformatics analysis revealed that C1 GALT1 was highly expressed in gastric cancer tissues,and its expression was closely correlated with the O-glycosylation and autophagy pathways.Mechanism investigation demonstrated that itraconazole downregulated C1 GALT1 protein expression,blocked O-glycosylation,and increased autophagy in gastric cancer cells.Additionally,C1 GALT1 overexpression attenuated the inhibitory effects of itraconazole on the invasion and migration of gastric cancer cells.Conclusion Itraconazole can inhibit gastric cancer cell invasion and metastasis by directly targeting C1 GALT1 and has anti-tumor activity in gastric cancer.

关 键 词：伊曲康唑 胃癌 C1GALT1 侵袭 转移 

分 类 号：R735.2[医药卫生—肿瘤]