Deng Yong, Yi Linglu, Lin Xuexia, Lin Ling, Li Haifang, Lin Jin-Ming
Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China; State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
Talanta. 2015 Nov 1;144:136-44. doi: 10.1016/j.talanta.2015.05.065. Epub 2015 May 27.
Bladder cancer (BC) cells spontaneously exfoliated in the urine of patients with BC. Detection of exfoliated tumor cells has clinical significance in cancer therapy because it would enable earlier non-invasive screening, diagnosis, or prognosis of BC. In this research, a method for analyzing genetic abnormalities of BC cells collected from urine samples was developed. Target BC cells were isolated by filtration. To find conditions that achieve high cell recovery, we investigated the effects of filter type, concentration of fixative, and flow rate. Cells captured on the filter membrane were completely retrieved within 15s. Selected genes for genomic analysis, mutated genes (FGFR3, TERT and HRAS) and methylated genes (ALX4, RALL3, MT1A, and RUNX3) were amplified by polymerase chain reaction (PCR), and subsequently, were identified by microchip electrophoresis (MCE). Analysis by MCE reduces the risk of contamination, sample consumption, and analysis time. Our developed approach is economical, effectively isolates cancer cells, and permits flexible molecular characterization, all of which make this approach a promising method for non-invasive BC detection.
膀胱癌（BC）患者尿液中会自发脱落癌细胞。检测脱落的肿瘤细胞在癌症治疗中具有临床意义，因为它能够实现对膀胱癌的早期非侵入性筛查、诊断或预后评估。在本研究中，开发了一种分析从尿液样本中收集的膀胱癌细胞基因异常的方法。通过过滤分离目标膀胱癌细胞。为了找到实现高细胞回收率的条件，我们研究了滤器类型、固定剂浓度和流速的影响。滤膜上捕获的细胞在15秒内被完全回收。用于基因组分析的选定基因，即突变基因（FGFR3、TERT和HRAS）和甲基化基因（ALX4、RALL3、MT1A和RUNX3）通过聚合酶链反应（PCR）进行扩增，随后通过微芯片电泳（MCE）进行鉴定。通过MCE进行分析可降低污染风险、样本消耗和分析时间。我们开发的方法经济实惠，能有效分离癌细胞，并允许进行灵活的分子特征分析，所有这些都使该方法成为一种有前景的非侵入性膀胱癌检测方法。
