Templeton Zachary S, Bachmann Michael H, Alluri Rajiv V, Maloney William J, Contag Christopher H, King Bonnie L
Department of Pediatrics, Stanford University School of Medicine.
Department of Orthopaedic Surgery, Stanford University School of Medicine.
J Vis Exp. 2015 Mar 15(97):52656. doi: 10.3791/52656.
Bone is the most common site of breast cancer metastasis. Although it is widely accepted that the microenvironment influences cancer cell behavior, little is known about breast cancer cell properties and behaviors within the native microenvironment of human bone tissue.We have developed approaches to track, quantify and modulate human breast cancer cells within the microenvironment of cultured human bone tissue fragments isolated from discarded femoral heads following total hip replacement surgeries. Using breast cancer cells engineered for luciferase and enhanced green fluorescent protein (EGFP) expression, we are able to reproducibly quantitate migration and proliferation patterns using bioluminescence imaging (BLI), track cell interactions within the bone fragments using fluorescence microscopy, and evaluate breast cells after colonization with flow cytometry. The key advantages of this model include: 1) a native, architecturally intact tissue microenvironment that includes relevant human cell types, and 2) direct access to the microenvironment, which facilitates rapid quantitative and qualitative monitoring and perturbation of breast and bone cell properties, behaviors and interactions. A primary limitation, at present, is the finite viability of the tissue fragments, which confines the window of study to short-term culture. Applications of the model system include studying the basic biology of breast cancer and other bone-seeking malignancies within the metastatic niche, and developing therapeutic strategies to effectively target breast cancer cells in bone tissues.
骨是乳腺癌转移最常见的部位。尽管人们普遍认为微环境会影响癌细胞的行为，但对于人骨组织天然微环境中乳腺癌细胞的特性和行为却知之甚少。我们已开发出一些方法，用于追踪、量化和调节从全髋关节置换手术后废弃的股骨头中分离出的培养人骨组织碎片微环境中的人乳腺癌细胞。利用经过基因工程改造以表达荧光素酶和增强型绿色荧光蛋白（EGFP）的乳腺癌细胞，我们能够使用生物发光成像（BLI）可重复地定量迁移和增殖模式，使用荧光显微镜追踪骨碎片内的细胞相互作用，并通过流式细胞术评估定植后的乳腺细胞。该模型的主要优点包括：1）天然的、结构完整的组织微环境，其中包括相关的人类细胞类型；2）可直接进入微环境，这便于对乳腺和骨细胞的特性、行为及相互作用进行快速的定量和定性监测与干扰。目前，该模型的一个主要局限性是组织碎片的有限活力，这将研究窗口限制在短期培养。该模型系统的应用包括研究转移微环境中乳腺癌和其他亲骨性恶性肿瘤的基础生物学，以及制定有效靶向骨组织中乳腺癌细胞的治疗策略。
