Background/Objectives:Liquid biopsy methods have gained prominence as minimally invasive tools to improve cancer treatment outcomes. Circulating tumor cells (CTCs) offer valuable insights into both primary and metastatic lesions. However, validating the CTC test results requires confirmation that the detected cells originate from cancer tissue. While studies have identified CTCs in colorectal cancer (CRC) patients using molecular markers, simultaneous validation of their cancer tissue origin remains unexplored.Methods:This study introduces a simple approach to detect adenomatous polyposis coli (APC) gene abnormalities alongside established CTC markers using a molecular imaging flow cytometer (MI-FCM). Given that APC gene abnormalities occur in 60–70% of CRC patients, their detection serves as strong evidence of cancer origin.Results:Our method achieved 92% concordance with DNA sequence analysis of tumor-derived cells. In a proof-of-concept study using 5 mL of whole blood from CRC patients, we observed a high frequency of cells exhibiting APC abnormalities, cytokeratin (CK), and vimentin (Vim) expression. Extending the study to 80 CRC patients across pathological stages I–IV confirmed CK and Vim as valid CTC markers. Three distinct cell populations were identified in blood: CK+/Vim−, CK+/Vim+, and CK−/Vim+. CTC number and frequency increased progressively with cancer stage.Conclusions:This is the first report demonstrating CK and Vim as effective markers for direct CTC detection in CRC patients. Our findings provide evidence-based validation of CTC markers, offering new insights and advancing approaches for patient care.
**背景/目的：** 液体活检作为一种微创工具，在改善癌症治疗效果方面日益受到重视。循环肿瘤细胞（CTCs）为原发性和转移性病灶提供了重要信息。然而，验证CTC检测结果需确认所检细胞是否源自癌组织。尽管已有研究通过分子标志物在结直肠癌（CRC）患者中鉴定出CTCs，但对其癌组织来源的同时验证尚未得到充分探索。 **方法：** 本研究提出一种简便方法，利用分子成像流式细胞仪（MI-FCM）检测腺瘤性息肉病基因（APC）异常，并结合已确立的CTC标志物。鉴于APC基因异常在60–70%的CRC患者中出现，其检测可作为癌源性的有力证据。 **结果：** 本方法与肿瘤来源细胞的DNA序列分析结果一致性达92%。在一项概念验证研究中，使用CRC患者5 mL全血，我们观察到高频率细胞呈现APC异常、细胞角蛋白（CK）及波形蛋白（Vim）表达。进一步对80例病理分期I–IV期CRC患者的研究证实，CK和Vim可作为有效的CTC标志物。血液中鉴定出三种细胞亚群：CK+/Vim−、CK+/Vim+和CK−/Vim+。CTC数量及频率随癌症分期的进展而逐步增加。 **结论：** 本研究首次报道CK和Vim可作为CRC患者直接检测CTCs的有效标志物。研究结果为CTC标志物提供了基于证据的验证，为患者诊疗提供了新见解并推动了相关方法的进展。
肿瘤(癌症)患者之家