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

	A20 overexpression under control of mouse osteocalcin promoter in MC3T3-E1 cells inhibited tumor necrosis factor-alpha-induced apoptosis
	Qin, YJ; Zhang, ZL; Yu, LY; He, JW; Hou, YN; Liu, TJ; Wu, JC; Wu, SH; Guo, LH
刊名	ACTA PHARMACOLOGICA SINICA
	2006
卷号	27 期号:9 页码:1231-1237
关键词	A20 osteoblast apoptosis tumor necrosis factor-alpha
通讯作者	Zhang, ZL (reprint author), Shanghai Jiao Tong Univ, Ctr Preventing & Treating Osteoporosis, Osteoporosis Res Unit, Shanghai Peoples Hosp 6, Shanghai 200233, Peoples R China.,ZZL2002@medmail.com.cn
英文摘要	Aim: To construct an A20 expression vector under the control of mouse osteocalcin promoter (OC-A20), and investigate osteoblastic MC3T3-E1 cell line, which stably overexpresses A20 protein prevented tumor necrosis factor (TNF)-alpha-induced apoptosis. Methods: OC-A20 vector was constructed by fusing a fragment of the mouse osteocalcin gene-2 promoter with human A20 complementary DNA. Then the mouse MC3T3-E1 cell line, stably transfected by A20, was established. The expression of A20 mRNA and A20 protein in the cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. To determine the specificity of A20 expression in osteoblast, the mouse osteoblastic MC3T3-E1 cell line and mouse embryo fibroblast NIH3T3 cell line were transiently transfected with OC-A20. The anti-apoptotic role 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), respectively. Results: Weak A20 expression was found in MC3T3E1 cells with the primers of mouse A20. A20 mRNA and A20 protein expression were identified in MC3T3-E1 cells transfected with OC-A20 using RT-PCR and Western blot analysis. Only A20 mRNA expression was found in MC3T3-E1 cell after MC3T3-E1 cells and NIH3T3 cells were transient transfected with OC-A20. A decrease obviously occurred in the rate of apoptosis in the OC-A20 group compared with the empty vector (pcDNA3) group by FACS (P < 0.001). A significant increase in TUNEL positive staining was found in the pcDNA group compared with OC-A20 group (P < 0.001). Simultaneously, similar effects were demonstrated in DNA gel electrophoresis analysis. Conclusion: We constructed an osteoblast-specific expression vector that expressed A20 protein in MC3T3-E1 cells and confirmed that A20 protects osteoblast against TNF-alpha-induced apoptosis.
学科主题	Chemistry; Pharmacology & Pharmacy
类目[WOS]	Chemistry, Multidisciplinary ; Pharmacology & Pharmacy
关键词[WOS]	NF-KAPPA-B ; ZINC-FINGER PROTEIN ; ENDOTHELIAL-CELLS ; IN-VIVO ; GENE ; DEATH ; MICE ; TNF ; ACTIVATION ; MECHANISMS
收录类别	SCI
语种	英语
WOS记录号	WOS:000240554800017
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1759]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Qin, YJ,Zhang, ZL,Yu, LY,et al. A20 overexpression under control of mouse osteocalcin promoter in MC3T3-E1 cells inhibited tumor necrosis factor-alpha-induced apoptosis[J]. ACTA PHARMACOLOGICA SINICA,2006,27(9):1231-1237.
APA	Qin, YJ.,Zhang, ZL.,Yu, LY.,He, JW.,Hou, YN.,...&Guo, LH.(2006).A20 overexpression under control of mouse osteocalcin promoter in MC3T3-E1 cells inhibited tumor necrosis factor-alpha-induced apoptosis.ACTA PHARMACOLOGICA SINICA,27(9),1231-1237.
MLA	Qin, YJ,et al."A20 overexpression under control of mouse osteocalcin promoter in MC3T3-E1 cells inhibited tumor necrosis factor-alpha-induced apoptosis".ACTA PHARMACOLOGICA SINICA 27.9(2006):1231-1237.