MiR-129-5p通过靶定胸苷酸合成酶调节结肠癌细胞对5-氟尿嘧啶的敏感性  

MiR-129-5p can increase colorectal cancer cell sensitivity to 5-Fu by targeting TYMS

作　　者：胡俊华[1] 喻琴[1] 杨艳果[1] 王琦[1] 卢光新[1] 

机构地区：[1]湖北医药学院附属人民医院消化内科,湖北十堰442000

出　　处：《临床内科杂志》2014年第10期709-711,共3页Journal of Clinical Internal Medicine

摘　　要：目的 研究miR-129-5p通过靶定胸苷酸合成酶（TYMS）调控结肠癌细胞对5-氟尿嘧啶（5-Fu）敏感性的分子机制.方法 构建结肠癌耐药细胞株LoVo/5-Fu,并用实时荧光定量聚合酶链反应（PCR）检测miR-129-5p的表达水平.流式细胞术和MTT毒性实验检测过表达miR-129-5p对LoVo/5-Fu凋亡和半数致死剂量（IC50）的影响.构建TYMS 3＇UTR区荧光素酶报告载体,验证miR-129-5p对TYMS的靶向调控作用.Western Blot检测转染miR-129-5p后LoVo/5-Fu细胞中TYMS的蛋白表达.在LoVo/5-Fu内转染miR-129-5P siRNA,抑制其表达,检测LoVo/5-Fu细胞的IC50和凋亡.结果 过表达miR-129-5p后,LoVo/5-Fu细胞用5-Fu处理后凋亡增加,IC50降低.荧光素酶活性检测表明miR-129-5p能够抑制TYMS的荧光素酶活性.在LoVo/5-Fu细胞中miR-129-5p与TYMS的表达呈负相关.敲减TYMS后,LoVo/5-Fu细胞用5-Fu处理后凋亡增加,IC50降低.结论 MiR-129-5p能够通过靶定TYMS而增加结肠癌细胞对5-Fu的敏感性.Objective To investigate whether miR-129-5p affects colorectal cancer cells sensitivity to 5-Fu.Methods First,we constructed 5-Fu resistance colorectal cancer cell line LoVo/5-Fu and tested the expression level of miR-129-5p by quantitative real-time PCR.Second,we detected apoptosis and IC50 of LoVo/5-Fu by MTT and flow cytometry after transfection of miR-129-5p mimic.Third,we cloned 3＇UTR of TYMS into luciferase reporter vector to verify whether miR-129-5p could target TYMS 3＇UTR.Fourth,we tested the expression level of TYMS in LoVo/5-Fu cell after transfection of miRNA-129-5p mimics by western blotting.Finally,we detected the drug resistance and apoptosis of LoVo/5-Fu cells after knockdown of TYMS by siRNA.Results LoVo/5-Fu showed high resistance to 5-Fu and over-expression of miR-129-5p decreased LoVo/5-Fu resistance to 5-Fu（P ＜0.05）.MiR-129-5p could target the 3,UTR of TYMS.Over-expression of miR-129-5p inhibited the protein level of TYMS in LoVo/5-Fu cell line.Knockdown of TYMS increased apoptosis level of LoVo/5-Fu and decreased IC50 of 5-Fu.Conclusion MiR-129-5p can increase colorectal cancer cell sensitivity to 5-Fu by targeting TYMS.

关 键 词：MiR-129-5p TYMS 结肠癌 5-Fu敏感性 

分 类 号：R735.35[医药卫生—肿瘤]