circ-BCAR3通过调控miR-145-5p/VEGFR-2信号通路对胃癌细胞增殖和侵袭的抑制作用  

Inhibition of gastric cancer cell proliferation and invasion by circ-BCAR3 through regulation of miR-145-5p/VEGFR-2 signaling pathway

作　　者：熊燕鹃 柳永 彭会琴 徐咏 郭婷 Xiong Yanjuan;Liu Yong;Peng Huiqin;Xu Yong;Guo Ting(Department of Gastroenterology,Huangshi Central Hospital(Affiliated Hospital of Hubei Institute of Technology),Huangshi 435000,China;Department of Urology,Huangshi Central Hospital(Affiliated Hospital of Hubei Institute of Technology),Huangshi 435000,China;Department of Gastrointestinal Oncology,Peking University Cancer Hospital,Beijing 100142,China)

机构地区：[1]黄石市中心医院(湖北理工学院附属医院)消化内科,黄石435000 [2]黄石市中心医院(湖北理工学院附属医院)泌尿外科,黄石435000 [3]北京大学肿瘤医院胃肠肿瘤科,北京100142

出　　处：《国际生物医学工程杂志》2024年第6期577-583,共7页International Journal of Biomedical Engineering

基　　金：国家自然科学基金(81972758)。

摘　　要：目的分析环状RNA-BCAR3(circ-BCAR3)在胃癌组织中的表达意义,探讨沉默circ-BCAR3对胃癌细胞增殖和侵袭的影响及分子机制。方法利用UCSC数据库分析胃癌组织及其癌旁正常组织中circ-BCAR3的表达,并采用Kaplan-Meier法分析circ-BCAR3表达与胃癌患者生存率的关系。将干扰小RNA(siRNA)-阴性对照(si-NC)与si-circ-BCAR3分别转染至胃癌BGC823细胞,记作si-NC组和si-circ-BCAR3组。采用实时荧光定量逆转录PCR(RT-qPCR)检测胃癌NCI-N87、AGS、BGC823、HS-746T、SGC7901细胞和永生化胃黏膜上皮GES-1细胞中circ-BCAR3基因表达。克隆形成实验和Transwell实验分别检测BGC823细胞的增殖和侵袭能力。利用Circinteractome在线网站预测circ-BCAR3与miR-145-5p的结合位点,双荧光素酶报告基因实验验证circ-BCAR3和miR-145-5p的靶向关系。利用UCSC数据库分析胃癌组织中circ-BCAR3和miR-145-5p表达的关系。免疫印迹法检测BGC823细胞中血管内皮生长因子受体-2(VEGFR-2)信号通路蛋白磷酸化VEGFR-2(p-VEGFR-2)、磷酸化磷脂酶C-λ(p-PLC-λ)、磷酸化细胞外信号调节激酶(p-Erk)和磷酸化P38(p-P38)的表达。结果胃癌组织中circ-BCAR3相对表达水平(8.04±2.34)明显高于癌旁正常组织(2.53±1.74),差异具有统计学意义(P<0.01)。circ-BCAR3低表达的胃癌患者生存率高于circ-BCAR3高表达的胃癌患者,差异具有统计学意义(P<0.05)。NCI-N87、HS-746T、AGS、BGC823、SGC7901细胞中circ-BCAR3相对表达水平(5.05±0.36、3.50±0.21、2.87±0.38、6.80±0.29、3.77±0.53)均高于GES-1细胞(1.02±0.21),差异均具有统计学意义(均P<0.01)。si-NC组和si-circ-BCAR3组克隆形成数分别为(126.70±21.78)、(44.89±8.33)个,侵袭细胞数分别为(53.81±5.92)、(24.91±4.56)个,差异具有统计学意义(P<0.01)。circ-BCAR3中的AACUGGA序列与miR-145-5p中的UUGACCU序列靶向结合。野生型circ-BCAR3(WT-circ-BCAR3)+miR-145-5p组荧光素酶活性(0.26±0.03)低于WT-circ-BCAR3+miR-NC组(1.00±0.05),�Objective To analyze the expression and significance of circ-BCAR3 in gastric cancer tissues,and to explore the effect and molecular mechanism of silencing circ-BCAR3 on the proliferation and invasion of gastric cancer cells.Methods The expression of circ-BCAR3 in gastric cancer tissues and adjacent normal tissues was analyzed using the UCSC database.The relationship between circ-BCAR3 expression and the survival rate of gastric cancer patients was analyzed by Kaplan-Meier method.Small interfering RNA(siRNA)-negative control(si-NC)and si-circ-BCAR3 were transfected into gastric cancer BGC823 cells,and recorded as si-circ-BCAR3 group and si-NC group,respectively.Real-time reverse transcription-PCR(RT-qPCR)was used to detect the expression of circ-BCAR3 gene in gastric cancer NCI-N87,AGS,BGC823,HS-746T,SGC7901 cells,and immortalized gastric mucosal epithelial GES-1 cells.The proliferation and invasion abilities of BGC823 cells were detected by clone formation assay and Transwell assay,respectively.The binding sites of circ-BCAR3 and miR-145-5p were predicted using the online Circinteractome platform,and their targeting relationship was verified by a dual luciferase reporter gene assay.The relationship between the expression of circ-BCAR3 and miR-145-5p in gastric cancer tissues was analyzed using the UCSC database.The expression levels of vascular endothelial growth factor receptor-2(VEGFR-2)signaling pathway proteins,including phosphorylated VEGFR-2(p-VEGFR-2),phosphorylated phospholipase C-λ(p-PLC-λ),phosphorylated extracellular signal-regulated kinase(p-Erk),and phosphorylated P38(p-P38),were detected in BGC823 cells using Western blotting analysis.Results The relative expression level of circ-BCAR3 in gastric cancer tissues(8.04±2.34)was significantly higher than that in adjacent normal tissues(2.53±1.74),and the difference was statistically significant(P<0.01).The survival rate of gastric cancer patients with low circ-BCAR3 expression was higher than that of gastric cancer patients with high circ-BCAR3 exp

关 键 词：胃癌 环状RNA-BCAR3 微小RNA-145-5p 细胞增殖 细胞侵袭 

分 类 号：R73[医药卫生—肿瘤]