Circulating tumor cells (CTCs) and circulating cancer-associated fibroblasts (cCAFs) have been individually considered strong indicators of cancer progression. However, technical limitations have prevented their simultaneous analysis in the context of CTC phenotypes different from epithelial. This study aimed to analyze CTCs and cCAFs simultaneously in the peripheral blood of 210 breast cancer patients using DAPI/pan-keratin (K)/vimentin (V)/alpha-SMA/CD29/CD45/CD31 immunofluorescent staining and novel technology—imaging flow cytometry (imFC). Single and clustered CTCs of different sizes and phenotypes (i.e., epithelial phenotype K+/V− and epithelial–mesenchymal transition (EMT)-related CTCs, such as K+/V+, K−/V+, and K−/V−) were detected in 27.6% of the samples and correlated with metastases. EMT-related CTCs interacted more frequently with normal cells and tended to occur in patients with tumors progressing during therapy, while cCAFs coincided with CTCs (mainly K+/V− and K−/V−) in seven (3.3%) patients and seemed to correlate with the presence of metastases, particularly visceral ones. This study emphasizes the advantages of imFC in the field of liquid biopsy and highlights the importance of multimarker-based analysis of different subpopulations and phenotypes of cancer progression-related cells, i.e., CTCs and cCAFs. The co-detection of CTCs and cCAFs might improve the identification of patients at higher risk of progression and their monitoring during therapy.
循环肿瘤细胞（CTCs）与循环癌症相关成纤维细胞（cCAFs）均被视为癌症进展的重要标志物。然而，由于技术限制，目前尚难以对非上皮表型的CTC表型进行同步分析。本研究采用DAPI/广谱角蛋白（K）/波形蛋白（V）/α-SMA/CD29/CD45/CD31免疫荧光染色及新型成像流式细胞术（imFC），对210例乳腺癌患者外周血中的CTCs与cCAFs进行同步检测。在27.6%的样本中检测到不同大小与表型的单个及簇状CTCs（包括上皮表型K+/V−及上皮-间质转化相关CTCs，如K+/V+、K−/V+和K−/V−），其存在与转移灶相关。上皮-间质转化相关CTCs与正常细胞相互作用更频繁，且多见于治疗期间肿瘤进展的患者；而cCAFs在7例（3.3%）患者中与CTCs（主要为K+/V−和K−/V−表型）共存，且似乎与转移灶（尤其是内脏转移）的存在相关。本研究凸显了imFC技术在液体活检领域的优势，并强调了基于多标志物分析癌症进展相关细胞（即CTCs与cCAFs）不同亚群及表型的重要性。CTCs与cCAFs的联合检测有望提升对高进展风险患者的识别能力，并优化治疗过程中的监测策略。
肿瘤(癌症)患者之家