微小RNA-410对细胞色素P450 3A5基因表达水平的调控的研究  

Micro RNA-410 is a promising regulating target for cytochrome P450 3A5 expression

作　　者：陆瑶[1] 戴海将 温改燕 唐青[1] 袁洪[1,2] 陈瑞芳[2] 

机构地区：[1]中南大学湘雅三医院临床药理中心,长沙410013 [2]中南大学湘雅三医院中心实验室,长沙410013

出　　处：《中国临床药理学杂志》2016年第24期2290-2292,共3页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81273594);国家自然科学基金面上项目(81470535;81370359);中南大学博士自主探索创新项目(2016zzts153)

摘　　要：目的研究微小RNA-410(miRNA)对细胞色素P450 3A5(CYP3A5)表达的特异调控。方法用Targetscan、miRanda和Pictar等靶基因预测软件预测可能作用CYP3A5基因的miRNA,并进行统计和筛选。用Hep G2细胞进行转染实验,miR-410 mimic转染组予以miR-410 mimic(类似物)转染48 h,miR-410inhibitor转染组予以miR-410 inhibitor(抑制药),空白对照组加入等量的细胞培养液转染48 h。用实时荧光聚合酶链式反应(RT-PCR)和Western-blot鉴定CYP3A5 mRNA和蛋白质的表达。结果 miR-410在很大程度与CYP3A5mRNA的3'-UTR结合。与空白对照组的CYP3A5蛋白质表达水平为2.61±0.53相比,miR-410 mimic转染组为1.47±0.13,miR-410 inhibitor转染组为3.50±0.65,差异均有统计学意义(P<0.01)。miR-410 mimic转染组和miR-410 inhibitor转染组的CYP3A5 mRNA的表达水平分别为1.81±0.11和1.82±0.41,差异无统计学意义(P>0.05)。结论 miR-410参与CYP3A5蛋白表达的调控,是一种转录后调控机制。Objective To clarify the regulation effect of micro RNA （miRNA） to cytochrome P450 3A5 （ CY3A5 ） expression. Methods Softwares （Targetsean, miranda and Pictar） were applied to predict the miRNA which could work on CYP3A5, and the results showed that micro RNA - 410 （ miR - 410） could bind with CYP3A5 mRNA 3＇- UTR; HepG2 was used to carry out the cell transfection experiments in 5 groups ： blank control group, miR - 410 mimic （ analogue ） group, miR -410 mimic negative control group, miR - 410 inhibitor （ inhibi- tors） group, miR-410 inhibitor negative control group. The real- time polymerase chain reaction and Western - blot were applied to detect expression level of CYP3A5 mRNA and protein after transfeetion for 48 hours. Results Compared with blank control group （ 2.61 ± 0. 53 ）, cYP3A5 protein expression level of miR -410 mimic transfection group increased significantly （ 1.47 ± 0. 13 ） , and the miR - 410 inhibitor transfection group decreased greatly （ 3.50 ± 0. 65 ）, with significant difference （P〈0.01）. Also there is no significant difference of CYF3A5 mRNA expression level between the two groups （ 1.81 ±0. 11 vs 1.82 -±0. 41, P 〉0. 05）. Conclusion miR- 410 can regulate the expression of CYP3A5 through transcriptional regulation mechanism.

关 键 词：微小RNA-410 细胞色素P450 3A5 基因多态性 

分 类 号：R97[医药卫生—药品]