Yamada-Hara Miki, Takahashi Naoki, Byun Ji Won, Zeng Liping, Wang Zhihe, Tanaka Arisachi, Malakoutikhah Zahra, Hayashi Tomoko, Webster Nicholas J G, Raz Eyal, Bertin Samuel
Division of Rheumatology, Autoimmunity, and Inflammation, Department of Medicine, University of California San Diego, La Jolla, California.
Division of Periodontology, Department of Oral Biological Science, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
Curr Protoc. 2024 Dec;4(12):e70071. doi: 10.1002/cpz1.70071.
Metastasis remains a leading cause of cancer-related mortality, yet its study has been constrained by the lack of reliable animal models that faithfully replicate this complex process. Syngeneic models for studying lung cancer metastasis are limited, with the Lewis lung carcinoma (LLC) model being the most commonly employed. The conventional LLC orthotopic model involves injecting LLC cells intravenously (i.v.) via the tail vein into syngeneic C57BL/6 mice. However, this model has significant drawbacks, such as tumor development in multiple anatomical sites, incomplete lung tumor penetrance, and challenges in monitoring lung tumor growth. This article highlights the advantages of using luciferase-expressing LLC cells combined with bioluminescence imaging (BLI) to quantify tumor progression in live animals. We demonstrate that both white- and black-furred C57BL/6 mice can be used for BLI. Finally, we propose that intranasal (i.n.) instillation of LLC cells offers a valuable alternative to the traditional i.v. tail vein injection method, particularly for its simplicity and improved reproducibility. Although the LLC i.n. model does not recapitulate the metastasis process via the blood vascular route, it is an effective model for studying tumor seeding within the lungs and is particularly useful for analyzing the impact of the lung microenvironment on tumor initiation and progression. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Lewis lung carcinoma intravenous injection method Support Protocol: In vivo bioluminescence imaging Basic Protocol 2: Lewis lung carcinoma intranasal instillation method.
转移仍然是癌症相关死亡的主要原因，然而，由于缺乏能够忠实复制这一复杂过程的可靠动物模型，其研究受到了限制。用于研究肺癌转移的同基因模型有限，其中Lewis肺癌（LLC）模型是最常用的。传统的LLC原位模型包括通过尾静脉将LLC细胞静脉内（i.v.）注射到同基因C57BL/6小鼠体内。然而，该模型存在显著缺点，如在多个解剖部位发生肿瘤、肺肿瘤穿透不完全以及监测肺肿瘤生长存在挑战。本文强调了使用表达荧光素酶的LLC细胞结合生物发光成像（BLI）来量化活体动物肿瘤进展的优势。我们证明白色和黑色被毛的C57BL/6小鼠均可用于BLI。最后，我们提出经鼻（i.n.）滴注LLC细胞为传统的静脉内尾静脉注射方法提供了一种有价值的替代方法，特别是因其操作简单且重现性提高。尽管LLC经鼻模型不能重现通过血管途径的转移过程，但它是研究肺内肿瘤播种的有效模型，对于分析肺微环境对肿瘤起始和进展的影响特别有用。© 2024 Wiley Periodicals LLC。基本方案1：Lewis肺癌静脉注射方法 支持方案：体内生物发光成像 基本方案2：Lewis肺癌经鼻滴注方法。
