Takahashi Yusuke, Ijiri Yuichi, Fujino Shiki, Elnaz Nakhaei, Kishimoto Ayuko, Shirai Kentaro, Iwanaga Shigeki, Yanagida Masatoshi, Bhagat Ali Asgar S, Miyoshi Norikatsu
Department of Central Research Laboratories, Sysmex Corporation, Kobe 651-2271, Japan.
Department of Gastroenterology, Central Clinical School, Monash University, Melbourne 3004, Australia.
Cancers (Basel). 2025 Jan 18;17(2):303. doi: 10.3390/cancers17020303.
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. 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. 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. 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.
液体活检方法作为改善癌症治疗效果的微创工具已备受关注。循环肿瘤细胞（CTC）为原发性和转移性病变提供了有价值的见解。然而，要验证CTC检测结果，需要确认检测到的细胞源自癌组织。虽然已有研究使用分子标志物在结直肠癌（CRC）患者中鉴定出CTC，但对其癌组织来源的同时验证仍未得到探索。本研究引入了一种简单的方法，使用分子成像流式细胞仪（MI-FCM），在既定的CTC标志物之外检测腺瘤性息肉病（APC）基因异常。鉴于60%-70%的CRC患者存在APC基因异常，对其进行检测可作为癌症起源的有力证据。我们的方法与肿瘤来源细胞的DNA序列分析达成了92%的一致性。在一项使用CRC患者5毫升全血的概念验证研究中，我们观察到出现APC异常、细胞角蛋白（CK）和波形蛋白（Vim）表达的细胞频率很高。将研究扩展至80名I-IV期的CRC患者，证实CK和Vim是有效的CTC标志物。在血液中鉴定出三种不同的细胞群体：CK+/Vim-、CK+/Vim+和CK-/Vim+。CTC数量和频率随癌症分期逐渐增加。这是首份证明CK和Vim作为CRC患者直接CTC检测有效标志物的报告。我们的研究结果为CTC标志物提供了基于证据的验证，为患者护理提供了新见解并推进了相关方法。
