miR-9-5p通过靶向FOXO1基因调控食管癌细胞增殖、侵袭和迁移  

miR-9-5p regulates proliferation, invasion and migration of esophageal cancer cells by targeting FOXO1 gene

作　　者：时军利[1] 王磊[1] 王春青[1] 李萍[1] 何培元[1] 李炳庆[1] SHI Junli;WANG Lei;WANG Chunqing;LI Ping;HE Peiyuan;LI Bingqing(Department of Gastroenterology,the Affiliated Hospital of Chengde Medical College,Chengde 067000,China)

机构地区：[1]承德医学院附属医院消化内科

出　　处：《胃肠病学和肝病学杂志》2019年第6期644-649,共6页Chinese Journal of Gastroenterology and Hepatology

基　　金：承德市科学技术研究与发展计划项目(201601A028)

摘　　要：目的 探讨miR-9-5p通过靶向叉头框转录因子O1(FOXO1)对食管癌Eca-109细胞体外增殖、侵袭和迁移能力的调控作用。方法在食管癌Eca-109细胞中分别转染miR-9-5p inhibitor和NC-inhibitor,分别设置为anti-miR-9-5p组和NC组,并设置未转染的细胞为Blank组。采用实时荧光定量PCR(qRT-PCR)技术检测各组细胞中miR-9-5p的表达水平,噻唑蓝(MTT)法、Transwell实验分别检测细胞增殖、侵袭和迁移能力。通过生物信息学在线软件预测miR-9-5p与FOXO1互补结合位点,双荧光素酶报告基因实验验证二者靶向关系。采用qRT-PCR和蛋白免疫印迹法(Western blotting)分析miR-9-5p对FOXO1的调控作用。结果 anti-miR-9-5p组Eca-109细胞中miR-9-5p的表达水平、 OD 值、侵袭和迁移细胞数均显著低于NC组和Blank组(P<0.05)。miR-9-5p与FOXO1存在互补结合位点,双荧光素酶报告基因实验分析结果显示,miR-9-5p和FOXO1能够靶向结合。anti-miR-9-5p组Eca-109细胞中FOXO1 mRNA和蛋白表达水平明显高于NC组和Blank组(P<0.05)。NC组和Blank组细胞中以上指标均无明显改变(P>0.05)。结论干扰miR-9-5p的表达可通过靶向FOXO1基因抑制食管癌Eca-109细胞的体外增殖、侵袭和迁移能力。Objective To investigate the regulation of miR-9-5Pon the proliferation, invasion and migration of esophageal cancer Eca-109 cells by targeting the forkhead transcription factor O1(FOXO1). Methods The miR-9-5Pinhibitor and NC-inhibitor were transfected into esophageal cancer Eca-109 cells, respectively. Anti-miR-9-5Pgroup and NC group were set respectively, and untransfected cells were set as control group. Real-time quantitative PCR(qRT-PCR) was used to detect the expression of miR-9-5Pin each group. MTT assay and Transwell assay were used to detect cell proliferation, invasion and migration. The complementary binding sites of miR-9-5Pand FOXO1 were predicted by bioinformatics online software, and the dual luciferase reporter gene experiments verified the targeting relationship. The regulation of miR-9-5Pon FOXO1 was analyzed by qRT-PCR and Western blotting. Results The expression level, OD value, invasion and migration number of miR-9-5Pin Eca-109 cells of anti-miR-9-5Pgroup were significantly lower than those of NC group and Blank group(P<0.05). The miR-9-5Phad a complementary binding site with FOXO1, and the results of the dual luciferase reporter gene assay showed that miR-9-5Pand FOXO1 could bind to each other. The expressions of FOXO1 mRNA and protein in Eca-109 cells of anti-miR-9-5Pgroup were significantly higher than those of NC group and Blank group(P<0.05). There were no significant changes in the above indexes in the NC group and Blank group(P>0.05). Conclusion Interfering with the expression of miR-9-5Pcan inhibit the proliferation, invasion and migration of esophageal cancer Eca-109 cells by targeting FOXO1 gene.

关 键 词：miR-9-5p FOXO1基因 食管癌细胞 增殖 侵袭 迁移 

分 类 号：R735.1[医药卫生—肿瘤]