YAP1与RNA分子m6A修饰配合对胃癌化疗敏感性的影响  

The Effect of YAP1 and RNA Molecular m6A Modification on Chemotherapy Sensitivity of Gastric Cancer

作　　者：郑黎明 甄生华 黄祖义 欧阳玉律 戴峰 ZHENG Liming;ZHEN Shenghua;HUANG Zuyi(Jianli People's Hospital,Hubei Jianli 433300,China)

机构地区：[1]湖北省监利市人民医院肿瘤内科,湖北监利433300

出　　处：《河北医学》2023年第11期1800-1805,共6页Hebei Medicine

基　　金：湖北省卫生健康委员会课题,(编号:WJ2019F084)。

摘　　要：目的:探讨Yes相关蛋白1(YAP1)与RNA分子N6-甲基腺苷(m6A)修饰交互调节对胃癌(GC)化疗敏感性的影响。方法:选择顺铂耐药的AGS/DDP及其亲本顺铂敏感的AGS作为细胞模型。通过免疫印迹分析细胞中甲基转移酶样3(METTL3)、YAP1蛋白表达,poly(A)RNA斑点印迹确定m6A修饰的程度。采用荧光素酶报告基因分析METTL3诱导的m6A修饰对YAP1翻译的影响,和Tunel检测顺铂治疗对METTL3或YAP1敲除的AGS/DDP细胞凋亡的影响。构建小鼠异种移植模型,考察METTL3敲除对体内顺铂的敏感性的影响。结果:与AGS细胞相比,m6A甲基转移酶METTL3的表达在AGS/DDP细胞(一种顺铂耐药细胞)中上调。此外,METTL3增加了AGS/DDP细胞中YAP1 mRNA的m6A修饰,从而提高了其翻译效率。相反,敲除AGS/DDP细胞中的METTL3会降低YAP1表达。METTL3敲除后的顺铂治疗诱导AGS/DDP细胞凋亡增多,并减少BALB/c裸鼠中AGS/DDP细胞衍生的肿瘤生长。结论:METTL3诱导的m6A修饰通过促进YAP1上调在GC获得性顺铂耐药中发挥重要作用,表明METTL3作为GC靶向化疗的潜在候选者。Objective:To investigate the effect of the interaction between Yes-associated protein 1(YAP1)and RNA molecule N6-methyladenosine(m6A)modification on the chemosensitivity of gastric cancer(GC).Methods:Cisplatin-resistant AGS/DDP cells and their cisplatin-sensitive parental AGS cells were used as cell models.The expression of methyltransferase-like 3(METTL3)and YAP1 proteins was analyzed using immunoblotting,and the extent of m6A modification was determined by Poly(A)RNA dot blotting.The influence of METTL3-induced m6A modification on YAP1 translation was assessed using a luciferase reporter gene.The impact of cisplatin treatment on apoptosis in METTL3-or YAP1-knockout AGS/DDP cells was examined by Tunel.A mouse xenograft model was established to study the effect of METTL3 knockout on cisplatin sensitivity in vivo.Results:The expression of m6A methyltransferase METTL3 was elevated in AGS/DDP cells,a cisplatin-resistant subline of GC cells,in comparison to the parental cells.METTL3 increased the m6A modification of YAP1 mRNA in AGS/DDP cells,enhancing its translation efficiency.Conversely,knockout of METTL3 in AGS/DDP cells reduced YAP1 expression and restored cisplatin sensitivity.Cisplatin treatment following METTL3 knockout induced apoptosis in AGS/DDP cells and reduced tumor growth derived from AGS/DDP cells in BALB/c nude mice.Conclusion:METTL3-induced m6A modification plays a significant role in acquired cisplatin resistance by promoting YAP1 upregulation,suggesting that METTL3 may be a potential candidate for targeted chemotherapy in GC.

关 键 词：Yes相关蛋白1 N6-甲基腺苷 胃癌 甲基转移酶样3 顺铂 

分 类 号：R735.2[医药卫生—肿瘤]