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The Effects of Human PEX, a C-terminal Hemopexin-like Domain of MMP-2, on the Growth and Metastasis of Human Breast Cancer BICR-H1 Cells |
ZHAO Wei, HAN Hai-bo, ZHANG Zhi-qian |
Key Laboratory of Carcinogenesis and Translational Research(Ministry of Education),School of Oncology,Peking University, Beijing Cancer Hospital & Institute,Beijing 100142, China |
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Abstract Objective:This study was designed to express human MMP-2 C-terminal hemopexin-like domain (PEX) in E. coli, and then to test its biological effects on angiogenesis, as well as on the growth and metastasis of human breast cancer BICR-H1 cells. Method:PEX was cloned into prokaryotic expression vector pET28a (+) and was induced to express in BL21(DE3)-pLys by IPTG. PEX-His fusion protein, which existed in the inclusion bodies, was denatured by urea method, then was renatured and purified though Ni-NTA agarose beads. The purified PEX-His was further characterized for its biological effects on angiogenesis inhibition, as well as on the suppression of the growth and metastases of human breast cancer BICR-H1 cells by employing the CAM assay. Results: Human PEX expressed in E. coli was able to inhibit the growth of human umbilical vein endothelial cells in a time and dose dependent manner, and could effectively inhibit the CAM angiogenesis. Furthermore, the growth and metastases of BICR-H1 cells were suppressed significantly by injection of 10μg PEX through the vein of chicken embryos, and they were completely disappeared when 30μg PEX was administrated. Conclusion: Human PEX expressed in E. coli is an effective angiongenesis inhibitor. It can inhibit the growth and metastases of BICR-H1 cells, and it might be useful in the treatment of angiongenesis associated diseases such as cancer.
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Received: 25 November 2010
Published: 01 April 2011
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