DDAH1-V3 transcript might act as miR-21 sponge to maintain balance of DDAH1-V1 in cultured HUVECs

作     者：KUANG Da-bin ZHOU Ji-peng YULin-yu ZENG Wen-jing XIAO Jian ZHANG Zan-lin CHEN Xiao-ping 

作者机构：Department of Clinical PharmacologyXiangya HospitalCentral South University Institute of Clinical PharmacologyCentral South UniversityHunan Key Laboratory of Pharmacogenetics Cooperative Innovation Center for Molecular Target New Drug StudyUniversity of South China Department of Cardiovascular MedicineXiangya HospitalCentral South University Department of PharmacyXiangya HospitalCentral South University 

出 版 物：《中国药理学与毒理学杂志》 (Chinese Journal of Pharmacology and Toxicology)

年 卷 期：2016年第30卷第10期

页      面：1007-1008页

核心收录：

学科分类：1002[医学-临床医学] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 10[医学] 

基　　金：The project supported by National Natural Science Foundation of China(81170091,81373489,81422052) Special Topic of the Major Subject of National Science and Technology(2013ZX09509-107) Provincial Natural Science Foundation of Hunan(13JJ1010) Funds for Hunan Education Department Program(12K006) 

主　　题：DDAH1 miR-21 microRNA sponge 

摘      要：OBJECTIVE To investigate whether micro RNA(mi RNA)mi R-21 regulates dimethylarginine dimethylaminohydrolase 1(DDAH1)expression through binding 3′-UTR regiondirectly in human umbilical venous endothelial cells(HUVECs)and to explore whether DDAH1-V2/V3 transcripts can function as micro RNA sponge,thereby modulating DDAH1-V1 *** The DDAH13′-UTR containing mi R-21 recognizing sequence was cloned into Pmir GLO dual-luciferase mi RNA target expression plasmid to construct PmirGLO-mi *** plasmid and mi R-21(at concentrations of 25,50,100 nmol·L-1,respectively)or negative control(100 nmol·L-1)were co-transfected into HUVECs,luciferase activity was detected at 24 *** were incubated with 2μg·m L-1actinomycin D for the indicated time after mi R-21(25 nmol.L-1)transfection,half-lives of DDAH1 m RNA were *** were transfected with Pmir GLO-mi R-21 alone or co-transfected with mi R-21 for 24 h,DDAH1 transcripts m RNA and DDAH1protein expression were *** Mi R-21decreased luciferase activity of Pmir GLO-mi R-21 in a dose-dependent manner(P0.05 for 25 nmol·L-1mi R-21,P0.01 for 50 nmol·L-1and 100 nmol·L-1mi R-21),and mi R-21 inhibitor increased reporter activity of PmirGLO-mi R-21 and m RNA expression of all DDAH1 three transcript variants significantly(P0.05,respectively).The degree of increase in endogenous DDAH1 m RNA expression by mi R-21 inhibitor was more obvious for *** of mi R-21 decreased m RNA expression and m RNA half-life time of all DDAH1 transcripts significantly(P0.05),and DDAH1-V2 displayed significantly decreased half-life time than DDAH1-V1and-V3 with or without mi R-21 transfection(P0.05,respectively).Mi R-21(100 nmol·L-1)decreased DDAH1protein expression significantly(P0.05),which was reversed by Pmir GLO-mi R-21 transfection(P0.05).Transfection of Pmir GLO-mi R-21 alone increased intracellular mi R-21 expression by approximately 5.6-fold,but only showed a trend of increase in DDAH1 pr