微小RNA-1-3p靶向调控中心粒蛋白F通过磷酸肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白通路影响胃癌恶性发展的研究  

Targeting microRNA-1-3p to regulate centroprotein F affects malignant development of gastric cancer through phosphoinositol 3 kinase/protein kinase B/mammalian target of rapamycin pathway

作　　者：胡毕文 何春华 Hu Biwen;He Chunhua(Department of Gastroenterology,Second Affiliated Hospital of Jiaxing University,Jiaxing 314000,China)

机构地区：[1]嘉兴大学附属第二医院胃肠外科,嘉兴314000

出　　处：《中华实验外科杂志》2024年第8期1730-1734,共5页Chinese Journal of Experimental Surgery

基　　金：嘉兴市科技计划项目(2022AY30030)。

摘　　要：目的探究微小RNA(miR)-1-3p靶向调控中心粒蛋白F(CENPF)通过磷酸肌醇3激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)通路影响胃癌(GC)恶性发展的机制。方法选取嘉兴大学附属第二医院胃肠外科收治的50例经手术治疗、且留有手术样本的胃癌肿瘤组织及邻近正常组织样本作为研究对象,采用实时实时荧光定量聚合酶链反应(qRT-PCR)检测胃癌组织及细胞中miR-1-3p的表达。细胞计数试剂盒-8(CCK-8)及克隆形成实验检测细胞增殖;Transwell检测细胞迁移及侵袭能力;流式细胞术检测细胞周期分布;蛋白质印迹法(Western blot)检测CENPF及PI3K/Akt/mTOR通路蛋白表达;双荧光素酶报告基因分析miR-1-3p和CENPF的靶向关系。组织之间的比较行配对设计或独立样本t检验,多组间的比较行单因素方差分析。结果从TCGA获取正常组织和癌症组织的miRNA表达谱,结果发现,胃癌组织及细胞系中(MGC803、AGS、MKN45、BGC-823)的miR-1-3p表达水平底于邻近正常组织及正常人胃上皮细胞(5.16±1.59、0.45±0.07、0.39±0.08、0.57±0.10、0.55±0.10比3.25±0.42、1.00±0.11,t=20.251,P<0.05);miR-1-3p对胃癌细胞增殖的影响:MGC803和AGS细胞转染miR-1-3p mimic 72 h的细胞活力明显低于mimic NC(0.49±0.10、0.51±0.06比0.86±0.10、0.85±0.11,t=19.205,P<0.05);MGC803及AGS细胞转染IN-miR-1-3p 72 h的细胞活力明显高于IN-NC(1.06±0.09、1.15±0.11比0.83±0.10、0.85±0.11,t=23.151,P<0.05)。miR-1-3p对胃癌细胞迁移侵袭的影响:MGC803和AGS细胞转染miR-1-3p mimic的迁移细胞数及侵袭细胞数明显低于mimic NC(82.37±9.53、85.41±8.29、73.58±9.87、79.63±8.22比138.94±15.63、140.75±16.84、114.76±10.45、121.33±9.46,t=31.012,P<0.05);MGC803及AGS细胞转染IN-miR-1-3p的迁移细胞数和侵袭细胞数明显高于IN-NC(192.45±16.52、203.71±17.63、158.69±11.59、162.78±14.25比125.96±13.55、129.76±11.67、105.45±10.63、109.52±9.48,t=37.135,P<0.05)。CENPF在胃�ObjectiveTo explore the effect of miR-1-3p on the malignant development of gastric cancer(GC)by targeting and regulating centroprotein F(CENPF)through phosphoinositol 3 kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)pathway.MethodsSelect 50 cases of gastric cancer tumor tissue and adjacent normal tissue samples treated with surgery and with surgical samples from the Department of Gastroenterology at the Second Affiliated Hospital of Jiaxing University as the research subjects,Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-1-3p in GC tissues and cells.Cell proliferation was detected by cell counting kit 8(CCK-8)and clonal formation assays.Transwell assay was used to detect cell migration and invasion.The cell cycle distribution was detected by flow cytometry.The protein expression of CENPF and PI3K/Akt/mTOR pathway was detected by Western blotting.Targeting relationship between miR-1-3p and CENPF was analyzed by dual luciferase reporter gene.Pairing design or independent sample t-test for comparison between organizations,one-way ANOVA for comparison between multiple groups.ResultsThe miRNA expression profiles of normal and cancer tissues obtained from TCGA showed that,the expression levels of miR-1-3p in gastric cancer tissues and cell lines(MGC803,AGS,MKN45,BGC-823)are lower than those in adjacent normal tissues and normal human gastric epithelial cells(5.16±1.59,0.45±0.07,0.39±0.08,0.57±0.10,0.55±0.10 vs.3.25±0.42,1.00±0.11,t=20.251,P<0.05);The effect of miR-1-3p on the proliferation of gastric cancer cells:The cell viability of MGC803 and AGS cells transfected with miR-1-3p mimic for 72 hours was significantly lower than that of mimic NC cells(0.49±0.10,0.51±0.06 vs.0.86±0.10,0.85±0.11,t=19.205,P<0.05);the cell viability of MGC803 and AGS cells transfected with IN-miR-1-3p for 72 hours was significantly higher than that of IN-NC cells(1.06±0.09,1.15±0.11 vs.0.83±0.10,0.85±0.11,t=23.151,P<0.05).The effect of miR

关 键 词：胃癌 微小RNA 中心粒蛋白F 磷酸肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白 细胞周期 

分 类 号：R735.2[医药卫生—肿瘤]