Full Text:   <3261>

CLC number: Q291; R34

On-line Access: 2012-05-04

Received: 2011-12-24

Revision Accepted: 2012-03-27

Crosschecked: 2012-04-18

Cited: 5

Clicked: 6016

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2012 Vol.13 No.5 P.356-363


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

Author(s):  Xiao-ying Wang, Zhong-hua Chen, Ru-yi Zhang, Sen-quan Liu, Zhu Mei, Ying-ying Yu, Xiong Zhang, Qiang Xia, Yue-min Ding

Affiliation(s):  Department of Gastroenterology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China; more

Corresponding email(s):   dingyuemin@zucc.edu.cn

Key Words:  Cell migration, Bone morphogenetic protein-2 (BMP-2), COS-7, Cofilin

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[J]. Journal of Zhejiang University Science B, 2012, 13(5): 356-363.

@article{title="Construction of a eukaryotic expression vector pEGFP-C1-BMP-2 and its effect on cell migration",
author="Xiao-ying Wang, Zhong-hua Chen, Ru-yi Zhang, Sen-quan Liu, Zhu Mei, Ying-ying Yu, Xiong Zhang, Qiang Xia, Yue-min Ding",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Construction of a eukaryotic expression vector pEGFP-C1-BMP-2 and its effect on cell migration
%A Xiao-ying Wang
%A Zhong-hua Chen
%A Ru-yi Zhang
%A Sen-quan Liu
%A Zhu Mei
%A Ying-ying Yu
%A Xiong Zhang
%A Qiang Xia
%A Yue-min Ding
%J Journal of Zhejiang University SCIENCE B
%V 13
%N 5
%P 356-363
%@ 1673-1581
%D 2012
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1100386

T1 - Construction of a eukaryotic expression vector pEGFP-C1-BMP-2 and its effect on cell migration
A1 - Xiao-ying Wang
A1 - Zhong-hua Chen
A1 - Ru-yi Zhang
A1 - Sen-quan Liu
A1 - Zhu Mei
A1 - Ying-ying Yu
A1 - Xiong Zhang
A1 - Qiang Xia
A1 - Yue-min Ding
J0 - Journal of Zhejiang University Science B
VL - 13
IS - 5
SP - 356
EP - 363
%@ 1673-1581
Y1 - 2012
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1100386

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.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Arnold, S.F., Tims, E., McGrath, B.E., 1999. Identification of bone morphogenetic proteins and their receptors in human breast cancer cell lines: importance of BMP2. Cytokine, 11(12):1031-1037.

[2]Blanco Calvo, M., Bolos Fernandez, V., Medina Villaamil, V., Aparicio Gallego, G., Diaz Prado, S., Grande Pulido, E., 2009. Biology of BMP signalling and cancer. Clin. Transl. Oncol., 11(3):126-137.

[3]Bobinac, D., Maric, I., Zoricic, S., Spanjol, J., Dordevic, G., Mustac, E., Fuckar, Z., 2005. Expression of bone morphogenetic proteins in human metastatic prostate and breast cancer. Croat. Med. J., 46(3):389-396.

[4]Fiedler, J., Roderer, G., Gunther, K.P., Brenner, R.E., 2002. BMP-2, BMP-4, and PDGF-bb stimulate chemotactic migration of primary human mesenchymal progenitor cells. J. Cell Biochem., 87(3):305-312.

[5]Foletta, V.C., Lim, M.A., Soosairajah, J., Kelly, A.P., Stanley, E.G., Shannon, M., He, W., Das, S., Massague, J., Bernard, O., 2003. Direct signaling by the BMP type II receptor via the cytoskeletal regulator LIMK1. J. Cell Biol., 162(6):1089-1098.

[6]Fong, Y.C., Li, T.M., Wu, C.M., Hsu, S.F., Kao, S.T., Chen, R.J., Lin, C.C., Liu, S.C., Wu, C.L., Tang, C.H., 2008. BMP-2 increases migration of human chondrosarcoma cells via PI3K/Akt pathway. J. Cell. Physiol., 217(3):846-855.

[7]Gamell, C., Osses, N., Bartrons, R., Ruckle, T., Camps, M., Rosa, J.L., Ventura, F., 2008. BMP2 induction of actin cytoskeleton reorganization and cell migration requires PI3-kinase and Cdc42 activity. J. Cell. Sci., 121(Pt 23):3960-3970.

[8]Gamell, C., Susperregui, A.G., Bernard, O., Rosa, J.L., Ventura, F., 2011. The p38/MK2/Hsp25 pathway is required for BMP-2-induced cell migration. PLoS One, 6(1):e16477.

[9]Goldstein, A.M., Brewer, K.C., Doyle, A.M., Nagy, N., Roberts, D.J., 2005. BMP signaling is necessary for neural crest cell migration and ganglion formation in the enteric nervous system. Mech. Dev., 122(6):821-833.

[10]Hocking, J.C., Hehr, C.L., Bertolesi, G., Funakoshi, H., Nakamura, T., McFarlane, S., 2009. LIMK1 acts downstream of BMP signaling in developing retinal ganglion cell axons but not dendrites. Dev. Biol., 330(2):273-285.

[11]Jiang, S., Zhang, S., Langenfeld, J., Lo, S.C., Rogers, M.B., 2008. Mycoplasma infection transforms normal lung cells and induces bone morphogenetic protein 2 expression by post-transcriptional mechanisms. J. Cell. Biochem., 104(2):580-594.

[12]Jin, Y., Tipoe, G.L., Liong, E.C., Lau, T.Y., Fung, P.C., Leung, K.M., 2001. Overexpression of BMP-2/4, -5 and BMPR-IA associated with malignancy of oral epithelium. Oral Oncol., 37(3):225-233.

[13]Kang, M.H., Oh, S.C., Lee, H.J., Kang, H.N., Kim, J.L., Kim, J.S., Yoo, Y.A., 2011. Metastatic function of BMP-2 in gastric cancer cells: the role of PI3K/AKT, MAPK, the NF-kappaB pathway, and MMP-9 expression. Exp. Cell Res., 317(12):1746-1762.

[14]Kraunz, K.S., Nelson, H.H., Liu, M., Wiencke, J.K., Kelsey, K.T., 2005. Interaction between the bone morphogenetic proteins and Ras/MAP-kinase signalling pathways in lung cancer. Br. J. Cancer, 93(8):949-952.

[15]Lai, T.H., Fong, Y.C., Fu, W.M., Yang, R.S., Tang, C.H., 2008. Osteoblasts-derived BMP-2 enhances the motility of prostate cancer cells via activation of integrins. Prostate, 68(12):1341-1353.

[16]Maiti, S.K., Singh, G.R., 1998. Bone morphogenetic proteins—novel regulators of bone formation. Indian J. Exp. Biol., 36(3):237-244.

[17]Matsui, S., Matsumoto, S., Adachi, R., Kusui, K., Hirayama, A., Watanabe, H., Ohashi, K., Mizuno, K., Yamaguchi, T., Kasahara, T., et al., 2002. LIM kinase 1 modulates opsonized zymosan-triggered activation of macrophage-like U937 cells. Possible involvement of phosphorylation of cofilin and reorganization of actin cytoskeleton. J. Biol. Chem., 277(1):544-549.

[18]Matthews, S.J., 2005. Biological activity of bone morphogenetic proteins (BMP’s). Injury, 36(S3):S34-S37.

[19]Sieber, C., Kopf, J., Hiepen, C., Knaus, P., 2009. Recent advances in BMP receptor signaling. Cytokine Growth Factor Rev., 20(5-6):343-355.

[20]Singh, A., Morris, R.J., 2010. The Yin and Yang of bone morphogenetic proteins in cancer. Cytokine Growth Factor Rev., 21(4):299-313.

[21]Sotobori, T., Ueda, T., Myoui, A., Yoshioka, K., Nakasaki, M., Yoshikawa, H., Itoh, K., 2006. Bone morphogenetic protein-2 promotes the haptotactic migration of murine osteoblastic and osteosarcoma cells by enhancing incorporation of integrin beta1 into lipid rafts. Exp. Cell Res., 312(19):3927-3938.

[22]Thawani, J.P., Wang, A.C., Than, K.D., Lin, C.Y., La Marca, F., Park, P., 2010. Bone morphogenetic proteins and cancer: review of the literature. Neurosurgery, 66(2):233-246.

[23]Urist, M.R., 1965. Bone: formation by autoinduction. Science, 150(3698):893-899.

[24]Vardouli, L., Moustakas, A., Stournaras, C., 2005. LIM-kinase 2 and cofilin phosphorylation mediate actin cytoskeleton reorganization induced by transforming growth factor-beta. J. Biol. Chem., 280(12):11448-11457.

[25]von Heijne, G., 1998. Life and death of a signal peptide. Nature, 396(6707):111, 113.

[26]Wozney, J.M., Rosen, V., Celeste, A.J., Mitsock, L.M., Whitters, M.J., Kriz, R.W., Hewick, R.M., Wang, E.A., 1988. Novel regulators of bone formation: molecular clones and activities. Science, 242(4885):1528-1534.

[27]Ye, L., Lewis-Russell, J.M., Kyanaston, H.G., Jiang, W.G., 2007. Bone morphogenetic proteins and their receptor signaling in prostate cancer. Histol. Histopathol., 22(10):1129-1147.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE