miR⁃21⁃5p靶向BNC2调控食管癌的恶性生物行为的研究  

Research on how miR⁃21⁃5p targets BNC2 to control the malignant biological activity of esophageal cancer

作　　者：邓雨函 阳梦 吴振华[1] 李卉 热则耶·麦麦提祖农 孙晓宏[1] DENG Yuhan;YANG Meng;WU Zhenhua;LI Hui;REZEYE•Maimaizunong;SUN Xiaohong(Xinjiang Medical University Affiliated Tumor Hospital,Urumqi 830054,China;Central Laboratory,Xinjiang Medical University,Urumqi 830054,China;School of Basic Medicine,Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学附属肿瘤医院,新疆乌鲁木齐830011 [2]新疆医科大学中心实验室,新疆乌鲁木齐830011 [3]新疆医科大学基础医学院,新疆乌鲁木齐830011

出　　处：《海南医学院学报》2024年第16期1236-1245,共10页Journal of Hainan Medical University

基　　金：新疆维吾尔自治区自然科学基金面上项目(2016D01C369,2021D01C397)。

摘　　要：目的:探讨miR-21-5p靶向肌成纤维细胞的身份转录因子(BNC2)调控食管癌(esophageal cancer,ESCA)细胞增殖、迁移、侵袭机制。方法:应用实时定量PCR(real-time quantitative polymerase chain reaction,qRT-PCR)方法检测食管癌组织中miR-21-5P和BNC2的相对表达量并用统计学分析其是否存在表达差异。用miR-21-5p mimics和miR-21-5P inhibitor和阴性对照(NC包括mimcs NC、inhibitor NC)分别转染ECA109、KYSE30两个食管癌细胞系后,进行细胞功能试验CCK8,Transwell等实验方法观察当miR-21-5p过表达或敲低时细胞的增殖、迁移和侵袭能力是否受到影响。通过生物信息学方法预测分析miR-21-5P的靶基因并通过双荧光素酶实验、qRT-PCR和Western blot实验验证miR-21-5p与靶向基因的负向调控关系。结果:qRT-PCR结果显示,相对于癌旁组织,BNC2在食管癌组织中的表达量显著性降低,而miR-21-5P则显著性升高,CCK8、Transwell实验结果显示,当miR-21-5P过表达时,细胞的增殖、迁移和侵袭能力增强,反之细胞功能受到抑制。也就是说,miR-21-5P在食管恶性肿瘤中起着促癌基因的功能作用。通过生物信息学方法预测miR-21-5P的靶基因为BNC2,双荧光素酶实验结果表明miR-21-5P与野生型BNC2′-UTR靶向结合,qRT-PCR和WB实验结果表示在食管癌细胞中,当miR-21-5P过表达时,BNC2的表达量降低,当miR-21-5P被敲低时,BNC2的表达量升高。这些结果显示miR-21-5P与BNC2存在靶向关系。结论:在食管癌中,miR-21-5p直接靶向BNC2,其调节作用可能从而导致癌症的增殖迁移和侵袭作用。Objective:To investigate the mechanism by which the identity transcription factor(BNC2)of myoblasts targeted by miR⁃21⁃5p in controling the proliferation,migration,and invasion of esophageal cancer(ESCA)cells.Methods:Real⁃time quantitative polymerase chain reaction(qRT⁃PCR)was used to assess the relative expression levels of miR⁃21⁃5P and BNC2 in esophageal cancer tissues.And any differences in expression were statistically analyzed.Following transfection of ECA109 and KYSE30 esophageal cancer cell lines with miR⁃21⁃5p mimics and miR⁃21⁃5P inhibitor,and a negative control(NC,including mimcs NC and inhibitor NC),respectively,the cell function test CCK8,and Transwell was used to investigate the impact of miR⁃21⁃5p downregulation or overexpression on cell migration,invasion,and proliferation.A bioinformatics technique was uti⁃lized to identify and investigate the target genes of miR⁃21⁃5P.Then,the dual luciferase assay,Western blot,and qRT⁃PCR as⁃say were employed to verify the negative regulatory relationship between miR⁃21⁃5p and the target genes.Result:In ESCA tumor tissues,it was possible to determine that BNC2 had reduced relative expression levels.and the relative expression levels of miR⁃21⁃5p were greater in tumor tissues by qRT⁃PCR detection.The outcomes of the CCK8 and Transwell studies demonstrated that overexpression of miR⁃21⁃5P increased cell invasion,migration,and proliferation.Cell invasion,migration,and proliferation were all decreased by knocking down miR⁃21⁃5P.Put differently,miR⁃21⁃5P functions as an oncogene in esophageal cancers.The bioinformatics technique predicted BNC2 to be the target gene of miR⁃21⁃5P.According to the results of the dual luciferase assay,miR⁃21⁃5P was targeted to wild⁃type BNC2′⁃UTR.Furthermore,miR⁃21⁃5p mimics were shown to be able to drastically lower the BNC2 gene level by western blot and qRT⁃PCR.On the other hand,a miR⁃21⁃5p inhibitor may dramatically raise BNC2 ex⁃pression in esophageal

关 键 词：食管癌 miR⁃21⁃5P BNC2 预后 肿瘤微环境 

分 类 号：R735.1[医药卫生—肿瘤]