GAS5作为miR-223-3p的ceRNA介导U87细胞的焦亡抑制脑胶质细胞的增殖,槲皮素可以增强GAS5的作用  

GAS5,as the ceRNA of miR-223-3p,mediates pyroptosis and inhibits the proliferation of brain glial cells U87,and quercetin can enhance its effect

作　　者：孙艳强 李虹良 陈康雪 李杰 孙中磊 Yanqiang Sun;Hongliang Li;Kangxue Chen;Jie Li;Zhonglei Sun(Department of Special Medical Center of Chinese People’s Armed Police Force,Emergency Department,Tianjin 300162,China;Chengdu Medical University,Chengdu 611137,Sichuan,China;Sichuan Taikang Hospital,Chengdu 610213,Sichuan,China)

机构地区：[1]中国人民武装警察部队特色医学中心急诊科,天津300162 [2]成都医学院,四川成都611137 [3]四川泰康医院,四川成都610213

出　　处：《Journal of Chinese Pharmaceutical Sciences》2024年第3期230-240,共11页中国药学（英文版）

基　　金：National Key Research and Development Project (Grant No.2016YFC1101503)。

摘　　要：本研究旨在探讨生长抑制特异性基因5(GAS5)作为miR-223-3p对脑胶质瘤生长的抑制作用及其可能机制。通过starbase工具预测GAS5与miR-223-3p结合情况,双荧光素酶报告实验和RNA免疫沉淀测定GAS5和miR-223-3p之间的结合关系。体外培养脑胶质细胞U87,分为对照组(control)、槲皮素组(quercetin)、GAS5抑制组(si-GAS5)和槲皮素+si-GAS5组(quercetin+si-GAS5)组,对比分析槲皮素对GAS5/miR-223-3p及焦亡蛋白(NLRP3、ASC、caspase-1、GSDMD和GSDME)的影响,以及U87细胞增殖能力的影响。结果显示,Starbase工具预测GAS5与miR-223-3p结合,双荧光素酶报告实验和RNA免疫沉淀测定揭示了GAS5和miR-223-3p之间的直接结合。槲皮素促进GAS5的表达,抑制miR-223-3p和焦亡相关蛋白(NLRP3、ASC、caspase-1、GSDMD和GSDME)的表达,抑制U87细胞的增殖和侵袭(P <0.05);下调GAS5, miR-223-3p和焦亡相关蛋白(NLRP3、ASC、caspase-1、GSDMD和GSDME)的表达增多,促进RC-4B/C细胞的增殖和侵袭(P<0.05);槲皮素可以逆转GAS5下调引起的miR-223-3p和焦亡相关蛋白(NLRP3、ASC、caspase-1、GSDMD和GSDME)的表达增加,抑制U87细胞的增殖和侵袭(P <0.05)。槲皮素抑制脑胶质瘤细胞U87的增殖,可能是通过促进GAS5表达竞争性抑制miR-223-3p,抑制焦亡信号途径实现的。This study explored the inhibitory effect of miR-223-3p on the proliferation of brain glioma cells and its potential mechanism in relation to growth arrest-specific transcripts(GAS5).We utilized the StarBase tool to predict the binding of GAS5 to miR-223-3p,further confirming the binding relationship between GAS5 and miR-223-3p through a dual luciferase reporter assay and RNA immunoprecipitation.In vitro cultured U87 brain glial cells were categorized into four groups:the control group,quercetin group,GAS5 inhibition group,and quercetin+si-GAS5 group.We analyzed and compared the impacts of quercetin on GAS5/miR-223-3p and pyroptotic proteins(NLRP3,ASC,caspase-1,GSDMD,and GSDME),as well as the proliferative capacity of U87 cells.StarBase’s predictions indicated that GAS5 bound to miR-223-3p.This was corroborated by the dual luciferase reporter assay and RNA immunoprecipitation assay,which demonstrated a direct binding between GAS5 and miR-223-3p.Quercetin was observed to enhance GAS5 expression,inhibit the expression of miR-223-3p and pyroptotic proteins(NLRP3,ASC,caspase-1,GSDMD,and GSDME),and suppress the proliferation and invasion of U87 cells(P<0.05).Downregulation of GAS5 expression correlated with an increase in the expression of miR-223-3p and pyroptotic proteins(NLRP3,ASC,caspase-1,GSDMD,and GSDME),promoting the proliferation and invasion of RC-4B/C cells(P<0.05).Quercetin was able to reverse the increase in miR-223-3p expression and pyroptotic proteins(NLRP3,ASC,caspase-1,GSDMD,and GSDME)induced by the downregulation of GAS5,thereby inhibiting the proliferation and invasion of U87 cells(P<0.05).In conclusion,quercetin might inhibit the proliferation of U87 brain glioma cells by promoting the expression of GAS5,which in turn competitively inhibited miR-223-3p and suppressed the pyroptotic signaling pathway.

关 键 词：脑胶质瘤 U87 GAS5 miR-223-3p 焦亡 

分 类 号：R961[医药卫生—药理学]