Reeves Kimberley J, Hurrell Jack E, Cecchini Marco, van der Pluijm Gabri, Down Jenny M, Eaton Colby L, Hamdy Freddie, Clement-Lacroix Philippe, Brown Nicola J
Microcirculation Research Group, Department of Oncology, CR-UK/YCR Sheffield Cancer Research Centre, Faculty of Medicine, Dentistry and Health, University of Sheffield, S10 2RX, United Kingdom; Bone Biology Group, Department of Human Metabolism, Medical School, University of Sheffield, Sheffield, S10 2RX, United Kingdom.
Int J Cancer. 2015 Apr 1;136(7):1731-40. doi: 10.1002/ijc.29165. Epub 2014 Sep 4.
Micrometastasis is a barrier to the development of effective cancer therapies for prostate cancer metastasis to bone. The mechanisms remain incompletely characterised, primarily due to an inability to adequately monitor the initial metastatic events in vivo. This study aimed to establish a new model, allowing the tracking of prostate cancer cells homing to bone, and furthermore, to evaluate the response of this approach to therapeutic modulation, using the integrin antagonist GLPG0187. A single murine metatarsal was engrafted into a dorsal skinfold chamber implanted on a SCID mouse. Fluorescently-labeled human prostate (PC3-GFP) or oral (SCC4-GFP) cancer cells were administered via intracardiac (i.c) injection, with simultaneous daily GLPG0187 or vehicle-control treatment (i.p. 100 mg/kg/day) for the experimental duration. Metatarsal recordings were taken every 48 h for up to 4 weeks. Tissue was harvested and processed for microCT, multiphoton analysis, histology and immunohistochemistry. Cell viability, proliferation and migration in vitro were also quantified following treatment with GLPG0187. Metatarsals rapidly revascularised by inosculation with the host vasculature (day 5-7). PC3-GFP cells adhered to the microvascular endothelium and/or metatarsal matrix 3 days after administration, with adhesion maintained for the experimental duration. GLPG0187 treatment significantly (p < 0.05) reduced PC3 cell number within the metatarsal in vivo and reduced migration (p < 0.05) and proliferation (p < 0.05) but not cell viability in vitro. This new model allows evaluation of the early events of tumour-cell homing and localisation to the bone microenvironment, in addition to determining responses to therapeutic interventions.
微转移是前列腺癌转移至骨的有效癌症治疗发展的一个障碍。其机制仍未完全明确，主要是由于无法在体内充分监测初始转移事件。本研究旨在建立一种新模型，用于追踪归巢至骨的前列腺癌细胞，此外，使用整合素拮抗剂GLPG0187评估该方法对治疗调节的反应。将一根小鼠跖骨植入植入SCID小鼠的背部皮褶小室中。通过心内注射给予荧光标记的人前列腺（PC3-GFP）或口腔（SCC4-GFP）癌细胞，在实验期间同时每日给予GLPG0187或载体对照治疗（腹腔注射100 mg/kg/天）。每48小时进行一次跖骨记录，持续4周。收集组织并进行显微CT、多光子分析、组织学和免疫组织化学处理。用GLPG0187处理后，还对体外细胞活力、增殖和迁移进行了定量分析。跖骨通过与宿主血管吻合在第5 - 7天迅速实现血管再生。给药后3天，PC3-GFP细胞粘附于微血管内皮和/或跖骨基质，并在实验期间保持粘附。GLPG0187治疗显著（p < 0.05）减少了体内跖骨内的PC3细胞数量，并降低了迁移（p < 0.05）和增殖（p < 0.05），但未影响体外细胞活力。这种新模型除了能确定对治疗干预的反应外，还允许评估肿瘤细胞归巢和定位于骨微环境的早期事件。
