A20 overexpression under control of mouse osteocalcin promoter in MC3T3-E1 cells inhibited tumor necrosis factor-alpha-induced apoptosis

作     者：Yue-juan QIN~2 Zhen-lin ZHANG~(2,4)Lu-yang YU~3 Jin-wei HE~2 Ya-nan HOU~3 Tian-jin LIU~3 Jia-cai WU~3 Song-hua WU~2 Li-he GUO~3 2 Center for Preventing and Treating Osteoporosis,Osteoporosis Research Unit,Shanghai Sixth People's Hospital,Shanghai Jiao Tong University,Shanghai 200233,China 3 Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Science,Chinese Academy of Sciences,Shanghai 200031,China 

作者机构：Center for Preventing and Treating Osteoporosis Osteoporosis Research Unit Shanghai Sixth People's Hospital Shanghai Jiao Tong University Shanghai China Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Science Chinese Academy of Sciences Shanghai China 

出 版 物：《Acta Pharmacologica Sinica》 (中国药理学报(英文版))

年 卷 期：2006年第27卷第9期

页      面：1231-1237页

核心收录：

学科分类：1007[医学-药学(可授医学、理学学位)] 1006[医学-中西医结合] 100706[医学-药理学] 100602[医学-中西医结合临床] 10[医学] 

基　　金：Project supported by the Natural Science Foundation of Shanghai(№03ZR14058) in part by National Natural Science Foundation of China(№30570891) 

主　　题：A20 osteoblast apoptosis tumor necrosis factor-alpha 

摘      要：Aim:To construct an A20 expression vector under the control of mouse osteocalcinpromoter(OC-A20),and investigate osteoblastic MC3T3-EI cell line,whichstably overexpresses A20 protein prevented tumor necrosis factor(TNF)-alpha-induced ***:OC-A20 vector was constructed by fusing a frag-ment of the mouse osteocalcin gene-2 promoter with human A20 *** the mouse MC3T3-E1 cell line,stably transfected by A20,*** expression of A20 mRNA and A20 protein in the cells weredetected by reverse transcription-polymerase chain reaction(RT-PCR)and West-ern blot analysis,*** determine the specificity of A20 expression inosteoblast,the mouse osteoblastic MC3T3-E1 cell line and mouse embryo fibro-blast NIH3T3 cell line were transiently transfected with *** anti-apoptoticrole of A20 in MC3T3-E1 cells was determined by Flow cytometric analysis(FACS),terminal dUTP nick endo-labeling(TUNEL)and DNA gel electrophoresis analysis(DNA Ladder),***:Weak A20 expression was found in MC3T3-E1 cells with the primers of mouse A20.A20 mRNA and A20 protein expressionwere identified in MC3T3-E1 cells transfected with OC-A20 using RT-PCR andWestern blot *** A20 mRNA expression was found in MC3T3-E1 cellafter MC3T3-E1 cells and NII-/3T3 cells were transient transfected with *** obviously occurred in the rate of apoptosis in the OC-A20 group com-pared with the empty vector(pcDNA3)group by FACS(P0.001).A significantincrease in TUNEL positive staining was found in the pcDNA group comparedwith OC-A20 group(P0.001).Simultaneously,similar effects were demonstratedin DNA gel electrophoresis ***:We constructed an osteoblast-specific expression vector that expressed A20 protein in MC3T3-E1 cells andconfirmed that A20 protects osteoblast against TNF-alpha-induced apoptosis.