Martinez-Marin Dalia, Jarvis Courtney, Nelius Thomas, Filleur Stéphanie
Department of Urology, Texas Tech University-Health Sciences Center, Lubbock, Texas 79430, USA.
Department of Immunology and Molecular Microbiology, Texas Tech University-Health Sciences Center, Lubbock, Texas 79430, USA.
Biol Methods Protoc. 2017 Mar 9;2(1):bpx002. doi: 10.1093/biomethods/bpx002. eCollection 2017 Jan.
Macrophages have been recognized as the main inflammatory component of the tumor microenvironment. Although often considered as beneficial for tumor growth and disease progression, tumor-associated macrophages have also been shown to be detrimental to the tumor depending on the tumor microenvironment. Therefore, understanding the molecular interactions between macrophages and tumor cells in relation to macrophages functional activities such as phagocytosis is critical for a better comprehension of their tumor-modulating action. Still, the characterization of these molecular mechanisms remains complicated due to the extraordinary complexity of the tumor microenvironment and the broad range of tumor-associated macrophage functions. Thus, there is an increasing demand for methodologies to study the role of cell-cell interactions in the tumor microenvironment. In the present study, we have developed live co-cultures of macrophages and human prostate tumor cells to assess the phagocytic activity of macrophages using a combination of Confocal and Nomarski Microscopy. Using this model, we have emphasized that this is a sensitive, measurable, and highly reproducible functional assay. We have also highlighted that this assay can be applied to multiple cancer cell types and used as a selection tool for a variety of different types of phagocytosis agonists. Finally, combining with other studies such as gain/loss of function or signaling studies remains possible. A better understanding of the interactions between tumor cells and macrophages may lead to the identification of new therapeutic targets against cancer.
巨噬细胞已被公认为肿瘤微环境的主要炎症成分。尽管肿瘤相关巨噬细胞通常被认为对肿瘤生长和疾病进展有益，但根据肿瘤微环境的不同，它们也可能对肿瘤产生不利影响。因此，了解巨噬细胞与肿瘤细胞之间与吞噬作用等巨噬细胞功能活动相关的分子相互作用，对于更好地理解它们的肿瘤调节作用至关重要。然而，由于肿瘤微环境的异常复杂性和肿瘤相关巨噬细胞功能的广泛多样性，这些分子机制的表征仍然很复杂。因此，对于研究肿瘤微环境中细胞间相互作用作用的方法的需求日益增加。在本研究中，我们建立了巨噬细胞与人类前列腺肿瘤细胞的活细胞共培养体系，结合共聚焦显微镜和微分干涉差显微镜来评估巨噬细胞的吞噬活性。利用该模型，我们强调这是一种灵敏、可测量且高度可重复的功能检测方法。我们还强调，该检测方法可应用于多种癌细胞类型，并可作为多种不同类型吞噬作用激动剂的筛选工具。最后，将其与其他研究（如功能获得/丧失或信号研究）相结合仍然是可行的。更好地理解肿瘤细胞与巨噬细胞之间的相互作用可能会导致识别出新的抗癌治疗靶点。
