通过将内皮抑素与BAX BH3死亡结构域融合来提高其治疗潜力。

Improving the therapeutic potential of endostatin by fusing it with the BAX BH3 death domain.

作者信息

Chura-Chambi R M, Bellini M H, Jacysyn J F, Andrade L N, Medina L P, Prieto-da-Silva A R B, Amarante-Mendes G P, Morganti L

机构信息

Centro de Biotecnologia, Instituto de Pesquisas Energéticas e Nucleares - IPEN - CNEN/SP, São Paulo, Brazil.

Laboratório de Investigação Médica LIM62, Faculdade de Medicina da, Universidade de São Paulo, São Paulo, Brazil.

出版信息

Cell Death Dis. 2014 Aug 14;5(8):e1371. doi: 10.1038/cddis.2014.309.

DOI:10.1038/cddis.2014.309

PMID:25118931

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4454297/

Abstract

Endostatin (ES) inhibits angiogenesis, reducing tumor growth in animal models. However, it has low therapeutic effect in human clinical trials. BAX is a member of the BCL-2 family of proteins; its proapoptotic (BH3) domain interacts with other members of the family in the cytoplasm, to induce apoptosis. Here, we fused the BAX BH3 domain with murine ES, to enhance ES potency. Endothelial cells specifically internalize the fusion protein ES-BAX. The presence of the BAX domain enhances endothelial cell death by apoptosis by 1.8-fold and diminishes microvessel outgrowth in the rat aortic ring assay by 6.5-fold. Daily injections of 15 μg of ES-BAX/g in tumor-bearing mice reduce tumor weight by 86.9% as compared with ES-treated animals. Co-immunoprecipitation assays confirmed that ES-BAX interacts with members of the BCL-2 family. Also, ES interacts with BCL-2, BCL-XL, and BAK in endothelial cell lysates, suggesting a potential new mechanism for the apoptosis induction by ES. The superiority of the ES-BAX antiangiogenic effect indicates that this fusion protein could be a promising therapeutic alternative to treat cancer.

摘要

内皮抑素（ES）可抑制血管生成，在动物模型中能减少肿瘤生长。然而，其在人体临床试验中的治疗效果不佳。BAX是BCL-2蛋白家族的成员之一；其促凋亡（BH3）结构域在细胞质中与该家族的其他成员相互作用，从而诱导细胞凋亡。在此，我们将BAX BH3结构域与鼠源ES融合，以增强ES的效能。内皮细胞可特异性内化融合蛋白ES-BAX。BAX结构域的存在使内皮细胞凋亡导致的细胞死亡增加了1.8倍，并在大鼠主动脉环试验中使微血管生长减少了6.5倍。在荷瘤小鼠中每日注射15μg ES-BAX/g，与接受ES治疗的动物相比，肿瘤重量减轻了86.9%。免疫共沉淀试验证实ES-BAX与BCL-2家族成员相互作用。此外，ES在内皮细胞裂解物中与BCL-2、BCL-XL和BAK相互作用，提示ES诱导细胞凋亡可能存在一种新机制。ES-BAX抗血管生成作用的优越性表明，这种融合蛋白可能是一种有前景的癌症治疗替代方案。