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| Construction of a eukaryotic expression vector pEGFP-C1-BMP-2 and its effect on cell migration |
| Xiao-ying Wang, Zhong-hua Chen, Ru-yi Zhang, Sen-quan Liu, Zhu Mei, Ying-ying Yu, Xiong Zhang, Qiang Xia, Yue-min Ding |
| Department of Gastroenterology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China; School of Medicine and Life Science, Zhejiang University City College, Hangzhou 310015, China; Department of Physiology, School of Medicine, Zhejiang University, Hangzhou 310058, China |
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Abstract 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.
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Received: 24 December 2011
Published: 04 May 2012
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Cite this article:
Xiao-ying Wang, Zhong-hua Chen, Ru-yi Zhang, Sen-quan Liu, Zhu Mei, Ying-ying Yu, Xiong Zhang, Qiang Xia, Yue-min Ding. Construction of a eukaryotic expression vector pEGFP-C1-BMP-2 and its effect on cell migration. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2012, 13(5): 356-363.
URL:
http://www.zjujournals.com/xueshu/zjus-b/10.1631/jzus.B1100386 OR http://www.zjujournals.com/xueshu/zjus-b/Y2012/V13/I5/356
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