Bao Q, Niess H, Djafarzadeh R, Zhao Y, Schwarz B, Angele M K, Jauch K-W, Nelson P J, Bruns C J
Department of Plastic and Reconstructive Surgery, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
Target Oncol. 2014 Sep;9(3):251-61. doi: 10.1007/s11523-013-0294-5. Epub 2013 Aug 10.
Fibrosarcomas show a high incidence of recurrence and general resistance to apoptosis. Limiting tumor regrowth and increasing their sensitivity to chemotherapy and apoptosis represent key issues in developing more effective treatments of these tumors. Tissue inhibitor of metalloproteinase 1 (TIMP-1) broadly blocks matrix metalloproteinase (MMP) activity and can moderate tumor growth and metastasis. We previously described generation of a recombinant fusion protein linking TIMP-1 to glycosylphophatidylinositol (GPI) anchor (TIMP-1-GPI) that efficiently directs the inhibitor to cell surfaces. In the present report, we examined the effect of TIMP-1-GPI treatment on fibrosarcoma biology. Exogenously applied TIMP-1-GPI efficiently incorporated into surface membranes of human HT1080 fibrosarcoma cells. It inhibited their proliferation, migration, suppressed cancer cell clone formation, and enhanced apoptosis. Doxorubicin, the standard chemotherapeutic drug for fibrosarcoma, was tested alone or in combination with TIMP-1-GPI. In parallel, the influence of treatment on HT1080 side population cells (exhibiting tumor stem cell-like characteristics) was investigated using Hoechst 33342 staining. The sequential combination of TIMP-1-GPI and doxorubicin showed more than additive effects on apoptosis, while TIMP-1-GPI treatment alone effectively decreased "stem-cell like" side population cells of HT1080. TIMP-1-GPI treatment was validated using HT1080 fibrosarcoma murine xenografts. Growing tumors treated with repeated local injections of TIMP-1-GPI showed dramatically inhibited fibrosarcoma growth and reduced angiogenesis. Intraoperative peritumoral application of GPI-anchored TIMP-1 as an adjuvant to surgery may help maintain tumor control by targeting microscopic residual fibrosarcoma cells and increasing their sensitivity to chemotherapy.
纤维肉瘤具有较高的复发率且普遍对凋亡有抗性。限制肿瘤再生长并提高其对化疗和凋亡的敏感性是开发更有效治疗这些肿瘤的关键问题。金属蛋白酶组织抑制剂1（TIMP-1）广泛阻断基质金属蛋白酶（MMP）的活性，并可调节肿瘤生长和转移。我们之前描述了一种将TIMP-1与糖基磷脂酰肌醇（GPI）锚定连接的重组融合蛋白（TIMP-1-GPI）的产生，该蛋白能有效地将抑制剂导向细胞表面。在本报告中，我们研究了TIMP-1-GPI处理对纤维肉瘤生物学特性的影响。外源性应用的TIMP-1-GPI能有效整合到人HT1080纤维肉瘤细胞的表面膜中。它抑制细胞增殖、迁移，抑制癌细胞克隆形成，并增强凋亡。阿霉素是纤维肉瘤的标准化疗药物，我们单独测试了它或与TIMP-1-GPI联合使用。同时，使用Hoechst 33342染色研究了处理对HT1080侧群细胞（表现出肿瘤干细胞样特征）的影响。TIMP-1-GPI和阿霉素的序贯联合对凋亡显示出超过相加的效应，而单独的TIMP-1-GPI处理有效地减少了HT1080的“干细胞样”侧群细胞。使用HT1080纤维肉瘤小鼠异种移植模型验证了TIMP-1-GPI处理。用TIMP-1-GPI重复局部注射治疗生长中的肿瘤，显示出纤维肉瘤生长显著受抑制且血管生成减少。术中在肿瘤周围应用GPI锚定的TIMP-1作为手术辅助手段，可能通过靶向微小残留纤维肉瘤细胞并提高它们对化疗的敏感性来帮助维持肿瘤控制。
