Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma
Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma
复制标题
口腔鳞状细胞癌原位小鼠模型中肿瘤细胞侵袭的多光子成像
DOI：
10.3791/2941
复制
发表时间：
2011-07-01
影响因子：
1.2
通讯作者：
Weed, Scott A.
中科院分区：
文献类型：
--
作者：
Ammer, Amanda Gatesman;Hayes, Karen E.;Weed, Scott A.
头颈部癌的局部区域侵袭与转移风险有关，并在设计和实施患者管理策略方面提出了困难的挑战。口腔癌的原位小鼠模型已经被开发，以促进影响侵袭的因素的研究，并作为评估抗肿瘤治疗的模型系统。在这些系统中，口腔组织内播散的肿瘤细胞的可视化通常通过常规组织学或体内生物发光方法进行。这些技术的主要缺点是固有的不能准确地可视化和量化从原发部位在三维中产生的早期肿瘤细胞侵袭。在这里，我们描述了一个协议，结合建立模型的舌鳞状细胞癌（SCOT）与双光子成像，允许多矢量可视化舌肿瘤扩散。将OSC-19头颈部肿瘤细胞系稳定地工程化以表达与mCherry荧光蛋白融合的F-肌动蛋白结合肽LifeAct（LifeAct-mCherry）。Fox 1（nu/nu）小鼠注射这些细胞可靠地形成肿瘤，使舌头可以通过离体应用双光子显微镜可视化。该技术允许原位可视化切除的舌中的肿瘤块和局部侵入细胞，而不破坏局部肿瘤微环境。此外，该系统允许通过计算侵入细胞从原发肿瘤部位移动的距离来量化肿瘤细胞侵入。总的来说，该程序提供了一个增强的模型系统，用于分析有助于SCOT侵袭和治疗性治疗的因素，以防止局部侵袭和远处转移扩散。这种方法也有可能最终与其他成像方式在体内设置相结合。
Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1(nu/nu) mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.