Cancer Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Orthopaedic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Mol Cancer Ther. 2015 Jul;14(7):1582-90. doi: 10.1158/1535-7163.MCT-14-0523. Epub 2015 May 1.
The aim of this study was to investigate the biologic role of the Rho kinase inhibitor fasudil in the vasculogenic mimicry (VM) of B16 mouse melanoma cells. It was previously reported that RhoA plays a critical role in angiogenesis by coordinating endothelial cell cytoskeleton remodeling and promoting endothelial cell motility. Although RhoA has been implicated in the regulation of angiogenesis, little has been described regarding its control of these tumor cell-lined channels. In this study, we established an in vitro model of VM using 3-dimensional cell culturing of mouse B16 melanoma cells and studied VM in vivo by transplanting B16 cells into C57/BL mice. Next, we explored the effect of RhoA and Rho-associated, coiled-coil containing protein kinase (ROCK) on VM formation using the Rho kinase inhibitor fasudil. We provide direct evidence that fasudil leads to reduced vascular-like channels in Matrigel. Additional experiments suggested that fasudil prevents both initial cellular architecture changes and cell migration in vitro. Finally, we provide in-depth evidence for the underlying mechanisms of fasudil-induced VM destruction using the Rho-GTPase agonist lysophosphatidic acid. In vivo studies revealed that fasudil reduced B16 melanoma cell xenograft tumor growth without causing significant toxicity in mice. Fasudil-treated tumors also displayed fewer VM channels. These results suggest that fasudil may be an emerging therapeutic option for targeting cancer VM.
本研究旨在探讨 Rho 激酶抑制剂法舒地尔在 B16 小鼠黑色素瘤细胞血管生成拟态（VM）中的生物学作用。先前的研究表明，RhoA 通过协调内皮细胞细胞骨架重塑和促进内皮细胞迁移，在血管生成中发挥关键作用。尽管 RhoA 已被认为参与了血管生成的调控，但关于其对这些肿瘤细胞排列的通道的控制作用，描述甚少。在本研究中，我们使用 3 维细胞培养的小鼠 B16 黑色素瘤细胞建立了体外 VM 模型，并通过将 B16 细胞移植到 C57/BL 小鼠中来研究体内 VM。接下来，我们使用 Rho 激酶抑制剂法舒地尔研究了 RhoA 和 Rho 相关卷曲螺旋蛋白激酶（ROCK）对 VM 形成的影响。我们提供了直接证据，表明法舒地尔导致 Matrigel 中血管样通道减少。进一步的实验表明，法舒地尔可防止体外细胞初始形态结构改变和细胞迁移。最后，我们使用 Rho-GTP 酶激动剂溶血磷脂酸，深入研究了法舒地尔诱导的 VM 破坏的潜在机制。体内研究表明，法舒地尔可减少 B16 黑色素瘤细胞异种移植肿瘤的生长，而对小鼠没有明显毒性。法舒地尔处理的肿瘤中 VM 通道也较少。这些结果表明，法舒地尔可能成为一种有前途的针对癌症 VM 的治疗选择。
