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
邓雨函 1阳梦 1吴振华 1李卉 2热则耶·麦麦提祖农 3孙晓宏1
作者信息
- 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
- 折叠
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、迁移和侵袭能力増强,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.
Abstract
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 cancer cells.These findings suggest a link of targeting between miR-21-5P and BNC2.Conclusion:In ES-CA,miR-21-5p targets BNC2,and its regulatory role may contribute to cancer proliferation,migration,and invasion.
关键词
食管癌/miR-21-5P/BNC2/预后/肿瘤微环境Key words
Tumor microenvironment/BNC2/MiR-21-5P/Esophageal cancer/Prognosis引用本文复制引用
基金项目
新疆维吾尔自治区自然科学基金面上项目(2016D01C369)
新疆维吾尔自治区自然科学基金面上项目(2021D01C397)
出版年
2024
国家科技期刊平台
NSTL
万方数据