Construction of a eukaryotic expression vector pEGFP-C1-BMP-2 and its effect on cell migration  

作　　者：Xiao-ying WANG Zhong-hua CHEN Ru-yi ZHANG Sen-quan LIU Zhu MEI Ying-ying YU Xiong ZHANG Qiang XIA Yue-min DING 

机构地区：[1]1Department of Gastroenterology,the Second Affiliated Hospital,School of Medicine,Zhejiang University,Hangzhou 310009,China [2]School of Medicine and Life Science,Zhejiang University City College,Hangzhou 310015,China [3]Department of Physiology,School of Medicine,Zhejiang University,Hangzhou 310058,China

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2012年第5期356-363,共8页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：Project supported by the National Natural Science Foundation of China(No.81000535);the Zhejiang Provincial Natural Science Foundation of China(No.Y2100508)

摘　　要：Objective: Bone morphogenetic proteins (BMPs) are known to play an important role in bone and cartilage development. Recent research has shown that BMPs may induce tumorigenesis and promote tumor to spread, but the molecular mechanisms have not been elucidated. The aim of the present study was to investigate the regulatory function of BMP-2 in the migration of COS-7 cells and the underlying mechanisms. Methods: Human BMP-2 genetic fragment was amplified and introduced into the pEGFP-C1 vector. After being confirmed by XhoI and BamHI digestion analyses and DNA sequencing, the recombinant pEGFP-C1-BMP-2 plasmid was transfected into COS-7 cells. The influence of BMP-2 on cell migration and cofilin activity was detected by cell scratch assay and Western blotting. Results: The recombinant pEGFP-C1-BMP-2 was effectively expressed in COS-7 cells. An increased phosphorylation of both LIMK1 and cofilin and an enhancement of cell migration were observed in cells with overexpression of BMP-2. Conclusions: A recombinant pEGFP-C1-BMP-2 vector was successfully constructed and overexpression of BMP-2 regulated the activities of the downstream molecules of the Rho GTPase signaling pathway, which might contribute to the enhancement of cell migration.Objective： Bone morphogenetic proteins （BMPs） are known to play an important role in bone and cartilage development. Recent research has shown that BMPs may induce tumorigenesis and promote tumor to spread, but the molecular mechanisms have not been elucidated. The aim of the present study was to investigate the regulatory function of BMP-2 in the migration of COS-7 cells and the underlying mechanisms. Methods： Human BMP-2 genetic fragment was amplified and introduced into the pEGFP-C1 vector. After being confirmed by XhoI and BamHI digestion analyses and DNA sequencing, the recombinant pEGFP-C1-BMP-2 plasmid was transfected into COS-7 cells. The influence of BMP-2 on cell migration and cofilin activity was detected by cell scratch assay and Western blotting. Results： The recombinant pEGFP-C1-BMP-2 was effectively expressed in COS-7 cells. An increased phosphorylation of both LIMK1 and cofilin and an enhancement of cell migration were observed in cells with overexpression of BMP-2. Conclusions： A recombinant pEGFP-C1-BMP-2 vector was successfully constructed and overexpression of BMP-2 regulated the activities of the downstream molecules of the Rho GTPase signaling pathway, which might contribute to the enhancement of cell migration.

关 键 词：Cell migration Bone morphogenetic protein-2 (BMP-2) COS-7 COFILIN 

分 类 号：Q291[生物学—细胞生物学] R34[医药卫生—基础医学]