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Neoplasma Vol.66, No.2, p.252-260, 2019 |
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Title: Fusion protein tTF-EG3287 induces occlusion of tumor vessels and impairs tumor growth in human colon cancer | ||
Author: G. Q. Qiu, X. Xie, B. Zhao, L. Z. Xu, Y. Q. Chen | ||
Abstract: The problems including narrow indications, low drug loading, and difficulty in intervention severely affect the clinical efficacy of anti-tumor embolization. Here, we designed a novel tTF-EG3287 protein consisting of the truncated tissue factor (tTF) fused with the bicyclic polypeptide which was encoded by exons 7 and 8 for accurate localization in the tumor vascular endothelial cells (EG3287). This study aims to explore its anti-cancer effect. Gene sequencing was used to verify the fusion gene and SDS-PAGE gel to confirm the optimal induction time and concentration of tTF-EG3287. Nickel affinity chromatography column was used to purify the fusion protein. Confocal microscopy was used to assess the target activity of tTF-EG3287 on colon cancer cells in vitro. Thrombelastography assay was used to identify the pro-coagulant activity of tTF-EG3287. In in vivo experiments, the specific localization of tTF-EG3287 in tumor tissues and the effect of tTF-EG3287 on tumor thrombosis were further detected by in vivo imaging and HE staining, respectively. The tTF-EG3287 fusion protein was efficiently purified by nickel-affinity chromatography column. Moreover, tTF-EG3287 fusion protein showed strong coagulation a ctivity and specific binding ability to the cell surface of colon cancer. In vivo, tTF-EG3287 stably and persistently accumulated in tumor tissues, and specifically induced mixed thrombus formation in tumor vessels, and then impaired tumor growth (tumor inhibition rate=79.2%, p |
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Keywords: colon cancer, tTF-EG3287, targeted therapy, vascular thrombosis, xenografts | ||
Published online: 19-Mar-2019 | ||
Year: 2019, Volume: 66, Issue: 2 | Page From: 252, Page To: 260 | |
doi:10.4149/neo_2018_180722N513 |
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