Despite advances in the detection and therapy of colorectal cancer (CRC) in recent years, CRC has remained a major challenge in clinical practice. Although alternative methods for modeling CRC have been developed, animal models of CRC remain helpful when analyzing molecular aspects of pathogenesis and are often used to perform preclinical in vivo studies of potential therapeutics. This protocol updates our protocol published in 2007, which provided an azoxymethane (AOM)-based setup for investigations into sporadic (Step 5A) and, when combined with dextran sodium sulfate (Step 5B), inflammation-associated tumor growth. This update also extends the applications beyond those of the original protocol by including an option in which AOM is serially applied to mice with p53 deficiency in the intestinal epithelium (Step 5C). In this model, the combination of p53 deficiency and AOM promotes tumor development, including growth of invasive cancers and lymph node metastasis. It also provides details on analysis of colorectal tumor growth and metastasis, including analysis of partial epithelial-to-mesenchymal transition, cell isolation and co-culture studies, high-resolution mini-endoscopy, light-sheet fluorescence microscopy and micro-CT imaging in mice. The target audience for our protocol is researchers who plan in vivo studies to address mechanisms influencing sporadic or inflammation-driven tumor development, including the analysis of local invasiveness and lymph node metastasis. It is suitable for preclinical in vivo testing of novel drugs and other interventional strategies for clinical translation, plus the evaluation of emerging imaging devices/modalities. It can be completed within 24 weeks (using Step 5A/C) or 10 weeks (using Step 5B).

尽管近年来结直肠癌 (CRC) 的检测和治疗取得了进展，但 CRC 仍然是临床实践中的主要挑战。尽管已经开发了用于建模 CRC 的替代方法，但在分析发病机制的分子方面时，CRC 的动物模型仍然很有帮助，并且通常用于对潜在治疗方法进行临床前体内研究。该协议更新了我们在 2007 年发布的协议，该协议提供了一种基于偶氮甲烷 (AOM) 的设置，用于调查散发性（步骤 5A）以及与葡聚糖硫酸钠（步骤 5B）结合时与炎症相关的肿瘤生长。此更新还通过包含一个选项将 AOM 连续应用于肠上皮中 p53 缺乏的小鼠（步骤 5C），从而将应用程序扩展到原始协议之外的应用程序。在这个模型中，p53 缺乏和 AOM 的结合促进了肿瘤的发展，包括浸润性癌症的生长和淋巴结转移。它还提供了结直肠肿瘤生长和转移分析的详细信息，包括部分上皮间质转化分析、细胞分离和共培养研究、高分辨率微型内窥镜检查、光片荧光显微镜和显微 CT 成像老鼠。我们协议的目标受众是计划进行体内研究以解决影响散发性或炎症驱动的肿瘤发展的机制的研究人员，包括分析局部侵袭性和淋巴结转移。它适用于新药的临床前体内测试和其他临床转化的介入策略，以及新兴成像设备/模式的评估。