miR-1827抑制肺腺癌A549细胞系迁移并调控c-Myc表达的研究  

MiR-1827 inhibits migration of lung adenocarcinoma A549 cell line and regulates c-Myc expression

作　　者：邓见青 初向阳 DENG Jianqing;CHU Xiangyang(Chinese PLA Medical School, Beijing 100853, China)

机构地区：[1]解放军医学院,北京100853

出　　处：《解放军医学院学报》2019年第2期172-177,共6页Academic Journal of Chinese PLA Medical School

摘　　要：目的探讨miR-1827对肺腺癌A549细胞细胞系迁移功能的影响以及对原癌基因c-Myc是否有调控作用。方法采用生物信息学方法对miR-1827与c-MYC基因mRNA 3'非翻译区(untranslated region,UTR)靶向配对关系进行预测。使用脂质体瞬时转染的方法将miR-1827类似物导入A549肺癌细胞系,使A549过表达miR-1827。Real-Time PCR验证转染后miR-1827以及c-Myc mRNA的表达量。Western blot法检测c-Myc蛋白表达量的变化。细胞划痕实验及Transwell细胞迁移实验检验miR-1827对A549迁移功能的影响。以上均以转染miR-NC序列的A549细胞作为对照。使用双萤光素酶报告基因实验对miR-1827与c-Myc是否存在直接调控关系及具体结合部位进行鉴定。使用GEO数据库的公开芯片数据对miR-1827与肺癌患者的预后之间的关系进行探索。结果在线生物信息学网站Targetscan以及DIANA-LAB显示miR-1827种子序列与c-Myc mRNA 3'UTR区存在完全配对序列。与对照组相比,miR-1827转染组miR-1827表达量显著上调,c-Myc基因mRNA以及蛋白水平均明显下降。双萤光素酶报告基因实验提示miR-1827直接通过与c-Myc mRNA 3'UTR配对结合下调c-Myc的表达。miR-1827可以显著抑制A549的迁移功能。芯片GSE63805显示miR-1827高表达组8年总生存率为56.25%,显著优于miR-1827低表达组的12.5%。结论 miR-1827能调控原癌基因c-Myc的表达并抑制A549细胞迁移。Objective To investigate the effect of miR-1827 on the migration of lung adenocarcinoma cell line A549 and whether it regulates the expression of proto-oncogene c-Myc. Methods The bioinformatics method was used to predict the paired sites between miR-1827 and c-Myc gene mRNA 3’ untranslated region(UTR). The miR-1827 mimics were transfected into the lung cancer cell line A549 using liposomes. Real-time PCR was performed to detect the expression of miR-1827 and c-Myc mRNA. The change of c-Myc protein expression was detected by western blot. Cell scratch assay and Transwell cell migration assay were performed to examine the effect of miR-1827 on the migration function of A549. A549 cells transfected with the miR-NC sequence were used as control. The dual luciferase reporter gene assay was used to identify whether miR-1827 directly bounded to c-Myc mRNA 3’UTR and speci?c binding sites. The association between miR-1827 expression and the prognosis of lung cancer patients was explored using published data from the GEO database. Results The online bioinformatics website Targetscan and DIANA-LAB showed that there was a mutual pairing sequence between the miR-1827 seed region and the c-Myc mRNA 3’ UTR region. Compared with miR-NC transfected group, the expression of miR-1827 was signi?cantly up-regulated, and the mRNA and protein levels of c-Myc gene decreased signi?cantly in the miR-1827 transfected group. The dual luciferase reporter assay suggested that miR-1827 downregulated c-Myc expression by pairing with c-Myc mRNA 3’UTR directly. MiR-1827 could significantly inhibit the migration function of A549. The data from chip GSE63805 showed that the 8-year survival rate of the miR-1827 high expression group was higher than that of the miR-1827 low expression group. Conclusion MiR-1827 can regulate the expression of proto-oncogene c-Myc and inhibit the migration of A549 cell line.

关 键 词：肺腺癌A549细胞系 微小RNA 原癌基因C-MYC 

分 类 号：R734.2[医药卫生—肿瘤]