Khoo Bee Luan, Bouquerel Charlotte, Durai Pradeep, Anil Sarannya, Goh Benjamin, Wu Bingcheng, Raman Lata, Mahendran Ratha, Thamboo Thomas, Chiong Edmund, Lim Chwee Teck
Department of Biomedical Engineering, City University of Hong Kong, 83 Tat Che e Avenue, Kowloon Tong, Hong Kong 999077, China.
Institut Pierre Gilles de Gennes, 75005 Paris, France.
Cancers (Basel). 2019 Aug 30;11(9):1274. doi: 10.3390/cancers11091274.
Bladder cancer (BC) is a disease that requires lifelong surveillance due to its high recurrence rate. An efficient method for the non-invasive rapid monitoring of patient prognosis and downstream phenotype characterization is warranted. Here, we develop an integrated procedure to detect aggressive mesenchymal exfoliated bladder cancer cells (EBCCs) from patients in a label-free manner. Using a combination of filtration and inertial focusing principles, the procedure allowed the focusing of EBCCs in a single stream-line for high-throughput separation from other urine components such as large squamous cells and blood cells using a microfluidic sorting device. Characterization of enriched cells can be completed within hours, suggesting a potential utility for real-time detection. We also demonstrate high efficiency of cancer cell recovery (93.3 ± 4.8%) and specific retrieval of various epithelial to mesenchymal transition (EMT) phenotype cell fractions from respective outlets of the microfluidic device. EMT is closely associated with metastasis, drug resistance and tumor-initiating potential. This procedure is validated with clinical samples, and further demonstrate the efficacy of bladder wash procedure to reduce EBCCs counts over time. Overall, the uniqueness of a rapid and non-invasive method permitting the separation of different EMT phenotypes shows high potential for clinical utility. We expect this approach will better facilitate the routine screening procedure in BC and greatly enhance personalized treatment.
膀胱癌（BC）因其高复发率而成为一种需要终身监测的疾病。因此，需要一种有效的方法来对患者预后进行无创快速监测以及对下游表型进行表征。在此，我们开发了一种综合程序，以无标记方式从患者中检测侵袭性间充质脱落膀胱癌细胞（EBCCs）。该程序结合了过滤和惯性聚焦原理，能够将EBCCs聚焦在单一流线中，以便使用微流控分选装置与其他尿液成分（如大的鳞状细胞和血细胞）进行高通量分离。富集细胞的表征可在数小时内完成，这表明其具有实时检测的潜在用途。我们还证明了癌细胞回收率高（93.3±4.8%），并且能够从微流控装置的各个出口特异性地分离出各种上皮-间充质转化（EMT）表型细胞组分。EMT与转移、耐药性和肿瘤起始潜能密切相关。该程序已通过临床样本验证，并进一步证明了膀胱冲洗程序随着时间推移减少EBCCs数量的有效性。总体而言，这种能够分离不同EMT表型的快速无创方法的独特性显示出很高的临床应用潜力。我们期望这种方法将更好地促进膀胱癌的常规筛查程序，并极大地加强个性化治疗。
