Division of Medical Oncology, Department of Internal Medicine, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, California, USA.
Cancer Res. 2010 Aug 15;70(16):6420-6. doi: 10.1158/0008-5472.CAN-10-0686. Epub 2010 Jul 27.
Circulating tumor cells (CTC) quantified in cancer patients' blood can predict disease outcome and response to therapy. However, the CTC analysis platforms commonly used cannot capture live CTCs and only apply to tumors of epithelial origin. To address these limitations, we have developed a novel cancer detection platform which measures telomerase activity from live CTCs captured on a parylene-C slot microfilter. Using a constant low-pressure delivery system, the new microfilter platform was capable of cell capture from 1 mL of whole blood in less than 5 minutes, achieving 90% capture efficiency, 90% cell viability, and 200-fold sample enrichment. Importantly, the captured cells retained normal morphology by scanning electron microscopy and could be readily manipulated, further analyzed, or expanded on- or off-filter. Telomerase activity--a well-recognized universal cancer marker--was reliably detected by quantitative PCR from as few as 25 cancer cells added into 7.5 mL of whole blood and captured on the microfilter. Moreover, significant telomerase activity elevation was also measured from patients' blood samples and from single cancer cells lifted off of the microfilter. Live CTC capture and analysis is fast and simple yet highly quantitative, versatile, and applicable to nearly all solid tumor types, making this a highly promising new strategy for cancer detection and characterization.
循环肿瘤细胞(CTC)在癌症患者血液中的定量可以预测疾病的结果和对治疗的反应。然而,目前常用的 CTC 分析平台无法捕获活的 CTC,并且仅适用于上皮来源的肿瘤。为了解决这些限制,我们开发了一种新的癌症检测平台,该平台通过在聚对二甲苯-C 插槽微滤器上捕获的活 CTC 来测量端粒酶活性。使用恒定的低压输送系统,新的微滤器平台能够在不到 5 分钟的时间内从 1 毫升全血中捕获细胞,实现 90%的捕获效率、90%的细胞活力和 200 倍的样品富集。重要的是,通过扫描电子显微镜可以看到捕获的细胞保留了正常形态,并且可以轻松操作、进一步分析或在滤器上或滤器下进行扩展。端粒酶活性——一种公认的通用癌症标志物——可以通过定量 PCR 从添加到 7.5 毫升全血中的 25 个癌细胞中可靠地检测到,并在微滤器上捕获。此外,还从患者的血液样本和从微滤器上取下的单个癌细胞中测量到显著的端粒酶活性升高。活 CTC 的捕获和分析快速、简单,同时具有高度定量、多功能和适用于几乎所有实体肿瘤类型的特点,这是一种极具前景的癌症检测和特征分析新策略。