癌細胞血管阻斷素（angiostatin）為內源性蛋白質，包含了血纖維蛋白溶解酶原（plasminogen）的前4個kringles。在活體實驗，癌細胞血管阻斷素會抑制小鼠的腫瘤生長，而且還可以抑制血管新生依賴型的腫瘤轉移。我們實驗室之前建構了一個K1-4/IgG嵌合蛋白質，它是由人類血纖維蛋白溶解酶原的kringle 1-4接上人類IgG的CH2-CH3所組成。在本篇研究中利用spinner flask及TideCell reactor兩種生物反應器系統生產K1-4/IgG蛋白質，研究發現，利用spinner flask 進行semi-perfusion culture的方法可以獲得最高的蛋白質產量（18.2mg/L），其產量比其它方法高出3～6倍。在活體實驗方面，將生產純化所獲得的K1-4/IgG蛋白質以及癌細胞血管阻斷素進行抑制小鼠血管新生及抗腫瘤之試驗，小鼠在接種小鼠膀胱癌細胞（MBT-2 cells）後，每天施以一定劑量的K1-4/IgG蛋白質或癌細胞血管阻斷素。由免疫組織染色以及抗腫瘤活體試驗可看出K1-4/IgG蛋白質以及癌細胞血管阻斷素不但可以減少腫瘤的微血管生成並可以明顯的抑制腫瘤細胞生長。此外，結合使用抗癌藥物doxorubicin與K1-4/IgG蛋白質比單獨使用doxorubicin更有效的抑制腫瘤生長。

Angiostatin is an internal proteolytically derived fragment of plasminogen. It spans the first four kringle domains. In in vivo studies, angiostatin could suppress tumor growth and angiogenesis-dependent tumor metastasis. A cell line that produces chimeric protein (K1-4/IgG) consisting of kringle 1-4 of human plasmonogen and CH2-CH3 of human IgG has been constructed in our laboratory. In this study, we used spinner flask and TideCell reactor as cell culture systems to produce K1-4/IgG protein. For this cell line, using semi-perfusion culture by spinner flask could gain the highest K1-4/IgG proteins production yield (18.2mg/L) as comparing to other methods. To evaluate the anti-angiogenesis effect of the K1-4/IgG protein in vivo, the murine tumor assay was used. After inoculation of MBT-2 cells, mice were treated with constant dosage of K1-4/IgG protein and angiostatin daily. In immunohistochemistry and in vivo anti-tumor test, K1-4/IgG protein and angiostatin could suppress angiogenesis around the tumor, and also could inhibit tumor growth. In addition, combination of using doxorubicin and K1-4/IgG protein could suppress tumor growth better than using doxorubicin alone.

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