Hsa-miR-411-3P对胃癌细胞作用功能及相关分子机制的研究  

Effect of Hsa-miR-411-3P on Gastric Cancer Cells and Related Mechanisms

作　　者：陈雪艳 张娜 陈娟 杨艳红[3] 张巨峰 CHEN Xue-yan;ZHANG Na;CHEN Juan;YANG Yan-hong;ZHANG Ju-feng(School of Pharmacy,Guangdong Pharmaceutical University,Guangzhou 510006,China;School of Life Science and Biopharmaceutics,Guangdong Pharmaceutical University,Guangzhou 510006,China;The First Affiliated Hospital,School of Clinical Medicine,Guangdong Pharmaceutical University,Guangzhou 510080,China)

机构地区：[1]广东药科大学药学院,广州510006 [2]广东药科大学生命科学与生物制药学院,广州510006 [3]广东药科大学附属第一医院(临床医学院),广州510080

出　　处：《中国生物工程杂志》2020年第4期1-9,共9页China Biotechnology

基　　金：广东省科技计划项目(2014A020212303);广东省中医药局科研项目(20182079);广州市越秀区科技计划(2018-WS-011)资助项目。

摘　　要：目的:研究Hsa-miR-411-3P对胃癌细胞功能作用及分子机制。方法:MTT检测Hsa-miR-411-3P对胃癌细胞SGC-7901、AGS增殖的影响;流式细胞术检测Hsa-miR-411-3P对胃癌细胞SGC-7901、AGS周期和凋亡的影响;RNAhybrid2.1.2、Miranda3.3a、TargetScan7.0预测Hsa-miR-411-3P靶基因,与m RNA测序结果联合分析,取交集基因认为是比较可靠的靶基因,进行GO、KEGG分析;Real-time PCR检测Hsa-miR-411-3P靶基因VAV3、ROCK2、PLD1、PTCH1的表达。结果:过表达Hsa-miR-411-3P抑制SGC-7901、AGS细胞增殖,促进凋亡,使SGC-7901细胞周期阻滞在S期,AGS细胞周期阻滞在G1期;软件预测和测序结果联合分析获得235个交集靶基因;Hsami R-411-3P靶基因分子功能集中在酶结合、蛋白质结合、转移酶活性等;生物学过程集中在代谢过程、细胞组件组装、解剖结构发展、细胞成分生物发生等(P<0.05);KEGG信号通路集中在胰岛素信号通路、cAMP信号通路、AMPK信号通路、FOXO信号通路等(P<0.05);VAV3、ROCK2、PLD1、PTCH1共同参与cAMP信号通路;过表达Hsa-miR-411-3P的SGC-7901、AGS细胞中,VAV3、ROCK2 m RNA表达量均减少,PLD1 m RNA表达量在SGC-7901细胞中增加,在AGS细胞中减少;PTCH1 m RNA表达量在SGC-7901细胞中减少,在AGS细胞中增加。结论:过表达Hsa-miR-411-3P将下调靶基因VAV3、ROCK2的表达,从而影响cAMP信号通路,抑制SGC-7901、AGS细胞增殖,促进凋亡,使SGC-7901细胞周期阻滞在S期,AGS细胞周期阻滞在G1期。Objective:To study the functional effects of Hsa-miR-411-3 P on gastric cancer cells and related molecular mechanisms.Method:The effect of Hsa-miR-411-3 P on the proliferation of gastric cancer cells SGC-7901 and AGS was detected by MTT;The effect of Hsa-miR-411-3 P on the cycle and apoptosis of gastric cancer cells SGC-7901、AGS was detected by flow cytometry;Three online tools:RNAhybrid2.1.2,Miranda3.3 a,TargetScan7.0 predicted Hsa-miR-411-3 P target genes,and combined with m RNA sequencing results,the intersection genes were considered reliable target genes,and then performed GO,KEGG analysis;Real-time PCR was used to detect the expression of Hsa-miR-411-3 P target genes VAV3,ROCK2,PLD1,and PTCH1.Result:Overexpression of Hsa-miR-411-3 P inhibited the proliferation of gastric cancer cells SGC-7901 and AGS,promoted the apoptosis of gastric cancer cells SGC-7901,AGS,arrested the cell cycle of SGC-7901 in S phase,and arrested AGS cell cycle in G1 phase.Three online tools:RNAhybrid2.1.2,Miranda3.3 a,TargetScan7.0,predicted Hsa-miR-411-3 P target genes,and combined with m RNA sequencing results,the intersection genes were considered to be reliable target genes,and a total of 235 intersection targets were obtained.Take 235 intersection target genes as target gene sets for subsequent analysis.GO and KEGG analysis of 235 cross-target genes found that their molecular function was concentrated in enzyme binding,protein binding,transferase activity,etc.Biological processes was concentrated in metabolic processes,assembly of cellular components,development of anatomical structures,and biogenesis of cellular components,etc.(P<0.05).KEGG signaling pathway was concentrated in insulin signaling pathway,cAMP signaling pathway,AMPK signaling pathway,FOXO signaling pathway,etc.(P<0.05).Insulin signaling pathway,AMPK signaling pathway,FOXO signaling pathway,and cAMP signaling pathway are all closely related to gastric cancer.The Hsa-miR-411-3 P target genes VAV3、ROCK2、PLD1 and PTCH1,which are highly correlated with gastric ca

关 键 词：胃癌 MIRNA Hsa-miR-411-3P 

分 类 号：R737.9[医药卫生—肿瘤]