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学科主题: 临床医学
题名:
Construction of recombinant adenovirus co-expression vector carrying the human transforming growth factor-beta 1 and vascular endothelial growth factor genes and its effect on anterior cruciate ligament fibroblasts
作者: Wei Xue-lei1; Lin Lin1; Hou Yu1; Fu Xin1; Zhang Ji-ying1; Mao Ze-bin2; Yu Chang-long1
关键词: adenovirus ; fibroblasts ; transforming growth factor-beta 1 ; vascular endothelial growth factor 165
刊名: CHINESE MEDICAL JOURNAL
发表日期: 2008-08-05
卷: 121, 期:15, 页:1426-1432
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Medicine, General & Internal
研究领域[WOS]: General & Internal Medicine
关键词[WOS]: MEDIAL COLLATERAL LIGAMENT ; HEALING PATELLAR LIGAMENT ; POLYMERASE CHAIN-REACTION ; RIBOSOME ENTRY SITES ; FACTOR-BETA ; IN-VITRO ; TENDON AUTOGRAFT ; RECONSTRUCTION ; RABBIT ; CELLS
英文摘要:

Background Remodeling of the anterior cruciate ligament (ACL) graft usually takes longer than expected. Gene therapy offers a radical different approach to remodeling of the graft. In this study, the internal ribosome entry site (IRES) sequence was used to construct a new recombinant adenovirus which permits co-expression of transforming growth factor-beta 1 (TGF beta 1) and vascular endothelial growth factor 165 (VEGF165) genes (named Ad-VEGF165-IRES-TGF beta 1). We investigated the effects of the new adenovirus on the migration of and matrix synthesis by ACL fibroblasts.

Methods Adenoviral vector containing TGF beta 1 and VEGF165 genes was constructed. ACL fibroblasts were obtained from New Zealand white rabbits. After ACL fibroblasts were exposed to Ad-VEGF165-IRES-TGF beta 1, the expression of VEGF165 and TGF beta 1 proteins were assessed by enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis. Bioassay of VEGF165 and TGF beta 1 proteins were assessed by Western blotting analysis. Proliferation and migration of ACL fibroblasts were assessed by in vitro wound closure assay. Gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers were assessed by real-time PCR.

Results The results showed the successful construction of a recombinant co-expression adenovirus vector containing TGF beta 1 and VEGF165 genes. Co-expression of TGF beta 1 and VEGF165 can induce relatively rapid and continuous proliferation of ACL fibroblasts and high gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers.

Conclusion Co-expression of TGF beta 1 and VEGF165 genes has more powerful and efficient effects on the migration of and matrix synthesis by ACL fibroblasts.

语种: 英语
项目资助者: Program on Gene Therapy in Sports Injuries ; State Sports General Administration of the People&prime ; s Republic of China
WOS记录号: WOS:000258421300016
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/57222
Appears in Collections:北京大学第三临床医学院_运动医学研究所_期刊论文

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作者单位: 1.Peking Univ, Inst Sports Med, Hosp 3, Beijing 100083, Peoples R China
2.Peking Univ, Dept Biochem & Mol Biol, Sch Basic Med Sci, Beijing 100083, Peoples R China

Recommended Citation:
Wei Xue-lei,Lin Lin,Hou Yu,et al. Construction of recombinant adenovirus co-expression vector carrying the human transforming growth factor-beta 1 and vascular endothelial growth factor genes and its effect on anterior cruciate ligament fibroblasts[J]. CHINESE MEDICAL JOURNAL,2008,121(15):1426-1432.
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