原花青素通过调节微小RNA-27a与微小RNA-451抑制多药耐药基因1 mRNA的表达  

Procyanidin inhibits multidruy resistance 1 mRNA expression via regulating microRNA-27a and microRNA-451

作　　者：张庆[1] 赵博欣[1] 王胜奇[1] 梁倩莹 杨富恒 王雅甜 蔡云[1] 李国锋[1] 

机构地区：[1]南方医科大学南方医院药学部,广州510515

出　　处：《国际药学研究杂志》2015年第4期501-506,共6页Journal of International Pharmaceutical Research

基　　金：南方医科大学南方医院院长基金资助项目(2012B012;2012C028);广东市科技计划资助项目(2014J4100144)

摘　　要：目的：研究原花青素对微小RNA-27a（miR-27a）与微小RNA-451（miR-451）在A2780/T细胞中表达的影响，及其与抑制多药耐药基因1（MDR1） mRNA表达的相关性。方法通过茎环法聚合酶链反应（stem-loop PCR），检测miR-27a与miR-451的表达水平，分别评价0~40μmol/L的原花青素刺激A2780/T细胞0~48 h后细胞内miR-27a与miR-451的表达水平。进一步通过转染miR-27a和miR-451的模拟或抑制片段，分别过表达或沉默相关的微小RNA，采用RT-PCR法评价原花青素对转染后细胞内MDR1 mRNA表达水平的影响。结果原花青素显著抑制了miR-27a与miR-451的表达水平，并且均具有浓度与时间依赖性。原花青素能够显著抑制过表达miR-27a与miR-451而上调的MDR1 mRNA水平，而对通过miR-27a与miR-451的沉默而下调的MDR1 mRNA水平无显著影响。结论原花青素能够通过抑制A2780/T细胞内miR-27a与miR-451的表达，从而抑制MDR1 mRNA的表达。Objective To investigate the effect of expression of microRNA-27a（miR-27a） and microRNA-451（miR-451） in A2780/T cells and its relativity to multidrug resistance （MDR）1 mRNA inhibition by procyanidin. Methods Stem-loop PCR method was performed to evaluate the expression of miR-27a and miR-451 in use of procyanidin （0-40μmol/L） in 0-48 h in A2780/T cells. Additionally, over-expressing or interfecting microRNAs by using mimics or inhibitor of miR-27a and miR-451, the expression of MDR1 mRNA was assessed by RT-PCR in cells exposing to procyanidin. Results The expression of miR-27a and miR-451 was significant inhibited by procyanidin in both time- and concentration-dependency. Over-expressed MDR1 mRNA associated with miR-27a or miR-451 mimics was blocked by procyanidin, whereas there was no effect on down-expressed MDR1 mRNA associated with miR-27a or miR-451 inhibitor by procyanidin. Conclusion Procyanidin inhibits MDR1 mRNA expression by inhibiting miR-27a and miR-451 expression in A2780/T cells.

关 键 词：原花青素 微小RNA 多药耐药基因1 茎环法聚合酶链反应 

分 类 号：R285[医药卫生—中药学]