转移是与癌症相关死亡的主要原因，迫切需要以转移为靶点的治疗方法。皮层细胞极性（CCP）是循环肿瘤细胞的一项促转移特性，影响它们从血管中离开并形成新的转移瘤，构成了预防转移的一个有希望的攻击点。然而，常规的荧光显微镜单细胞观察和CCP的手动计量非常耗时，并且不适合筛选调控因子。在本研究中，我们开发了一种基于成像流式细胞术的高通量筛选 CCP 影响因子的方法，该方法借助人工智能支持的分析方法，高度可重复、准确，并且比手动计量快得多。此外，该方法灵活性强，可以针对其他细胞参数进行调整。通过在黑色素瘤细胞中使用涉及极性、细胞骨架或细胞膜的药物进行小规模试验，我们证明了我们的工作流程提供了一个简单高效的方法，可以筛选悬浮细胞中影响 CCP 的因子，并揭示了这些药物在该细胞系统中的特定功能。本文介绍的方法和工作流程将有助于开展大规模研究，以揭示在转移过程中控制 CCP 的新型细胞内和系统性因子。© 2023 The Authors. Journal of Cellular Biochemistry published by Wiley Periodicals LLC.

Metastasis is the main cause of cancer-related death and therapies specifically targeting metastasis are highly needed. Cortical cell polarity (CCP) is a prometastatic property of circulating tumor cells affecting their ability to exit blood vessels and form new metastases that constitute a promising point of attack to prevent metastasis. However, conventional fluorescence microscopy on single cells and manual quantification of CCP are time-consuming and unsuitable for screening regulators. In this study, we developed an imaging flow cytometry-based method for high-throughput screening of factors affecting CCP in melanoma cells. The artificial intelligence-supported analysis method we developed is highly reproducible, accurate, and orders of magnitude faster than manual quantification. Additionally, this method is flexible and can be adapted to include additional cellular parameters. In a small-scale pilot experiment using polarity-, cytoskeleton-, or membrane-affecting drugs, we demonstrate that our workflow provides a straightforward and efficient approach for screening factors affecting CCP in cells in suspension and provide insights into the specific function of these drugs in this cellular system. The method and workflow presented here will facilitate large-scale studies to reveal novel cell-intrinsic as well as systemic factors controlling CCP during metastasis.© 2023 The Authors. Journal of Cellular Biochemistry published by Wiley Periodicals LLC.