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.
Abstract
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.
维普
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