miR-181a-2-3p提高头颈恶性肿瘤对5-氟尿嘧啶敏感性机制的研究  

Study of mechanism of miR-181a-2-3p enhancing the sensitivity of head and neck malignant tumors to 5-fluorouracil

作　　者：李增宏 廖烈强 何子键 黎景佳[1] 朱肇峰[1] 陈伟雄[1] LI Zenghong;LIAO Lieqiang;HE Zijian;LI Jingjia;ZHU Zhaofeng;CHEN Weixiong(Otolaryngology,Head and Neck Surgery Department,the First People's Hospital of Foshan,Foshan 528000,China)

机构地区：[1]广东省佛山市第一人民医院耳鼻咽喉头颈外科,广东佛山528000

出　　处：《临床医学研究与实践》2021年第22期22-27,共6页Clinical Research and Practice

基　　金：佛山市“登峰计划”科研项目(No.2019C004)。

摘　　要：目的筛查与5-氟尿嘧啶(5-FU)治疗头颈恶性肿瘤相关的微小RNA(microRNA),并对其所扮演的角色进行初步探讨。方法利用基因芯片对5-FU处理后的头颈恶性肿瘤细胞系中异常表达的microRNA进行分析,利用RT-PCR进行验证。预测microRNA的靶基因并利用RT-PCR及双荧光素酶报告基因实验进行验证。结果高通量检测提示,miR-29b-1-3p、miR-181a-2-3p、miR-486-3p的表达量同时在经5-FU治疗后的HN21B、HONE1、SNU899中上调,仅有miR-181a-2-3p的表达量在3个细胞系中随着5-FU的剂量增大均依次上调。上调细胞的miR-181a-2-3p表达量,3个细胞系5-FU的10%、30%、50%抑制浓度相较于空白对照组显著降低(P<0.05)。共发现miR-181a-2-3p的12个候选靶基因,通过RT-PCR验证及靶基因双荧光素酶报告基因实验发现,CITED2为miR-181a-2-3p在头颈恶性肿瘤中的靶基因,且随5-FU剂量增加显著下调。结论 miR-181a-2-3p/CITED2信号通路很可能参与调控头颈恶性肿瘤对于5-FU的敏感性。Objective To screen microRNA related to 5-fluorouracil(5-FU)in the treatment of head and neck malignant tumor,and to discuss their roles preliminarily.Methods Abnormal expression of microRNA in head and neck malignant tumors cell lines treated with 5-FU was analyzed by gene microarray,and verified by RT-PCR.The target genes of microRNA were predicted and verified by RT-PCR and dual luciferase reporter gene assay.Results High-throughput detection indicated that the expression levels of miR-29b-1-3p,miR-181a-2-3p and miR-486-3p up-regulated in HN21B,HONE1 and SNU899 after 5-FU treatment.Only the expression level of miR-181a-2-3p significantly up-regulated with the increase of 5-FU dose in all the three cell lines.When the expression of miR-181a-2-3p up-regulated,the 10%,30%and 50%inhibitory concentrations of 5-FU in three cell lines were significantly lower than those in the blank control group(P<0.05).A total of 12 candidate target genes of miR-181a-2-3p were found.Through RT-PCR verification and double luciferase reporter gene assay of target genes,it was found that CITED2 was the target gene of miR-181a-2-3p in head and neck malignant tumors,which was significantly down-regulated with the increase of 5-FU dose.Conclusion MiR-181a-2-3p/CITED2 signaling pathway may be involved in regulating the sensitivity of head and neck malignant tumors to 5-FU.

关 键 词：头颈恶性肿瘤 5-氟尿嘧啶 miR-181a-2-3p CITED2 

分 类 号：R967[医药卫生—药理学]