Cao Chengsong, Wang Qun, Li Qing, Zhao Qingqing, Wang Jun Qi, Liu Yong
J Biomed Nanotechnol. 2019 Mar 1;15(3):612-620. doi: 10.1166/jbn.2019.2706.
Real-time detection of circulating tumor cell (CTC) markers that are constantly changing and renewing during disease progression is of great significance for the timely regimen switch or individualized target therapy. The abnormally expressed special AT-rich sequence binding protein 1 (SATB1), a nuclear matrix attachment region binding protein, in various tumors, promotes the growth and metastasis of tumor cells by regulating gene expression. In this paper, a CTC detection system for prostate cancer (PCa) was developed on the basis of epithelial cell adhesion molecule (EpCAM)-targeted immunomagnetic separation and CK-FITC and SATB-1-APC immunofluorescence assay, and the recovery rate of tumor cells in PBS and simulated whole blood by this system was detected. Subsequently, we isolated, identified, and counted SATB-1 ositive CTCs in the peripheral blood and urine samples of 60 tumor-bearing nude mice, 5 healthy volunteers and 13 PCa patients. Combined with the clinicopathological factors, the clinical value of the system was analyzed, and the possibility of SATB-1-positive CTCs in the diagnosis of PCa was evaluated. The results showed that the CTC sorting and identification system for prostate cancer constructed in this study had a recovery rate of more than 85% for CTC in PBS, urine and blood simulation samples. The expression level of SATB-1 was different in different PCa cell lines, which was relatively high in the highly invasive PCa DU-145 cell line. The expression of SATB-1 in CTCs in the blood samples of PCa patients with different clinical characteristics and in the urine samples of a few PCa patients with bone metastases were different, and the detection sensitivity of peripheral blood was higher than that of urine. This study has important clinical reference value for the early diagnosis of PCa and the evaluation of bone metastasis based on the CTC counting and the SATB-1 expression in CTCs.
实时检测在疾病进展过程中不断变化和更新的循环肿瘤细胞（CTC）标志物，对于及时调整治疗方案或进行个体化靶向治疗具有重要意义。特殊富含AT序列结合蛋白1（SATB1）是一种核基质附着区域结合蛋白，在各种肿瘤中异常表达，通过调节基因表达促进肿瘤细胞的生长和转移。本文基于上皮细胞粘附分子（EpCAM）靶向免疫磁珠分离以及细胞角蛋白-异硫氰酸荧光素（CK-FITC）和SATB-1-别藻蓝蛋白（SATB-1-APC）免疫荧光检测，开发了一种用于前列腺癌（PCa）的CTC检测系统，并检测了该系统对PBS和模拟全血中肿瘤细胞的回收率。随后，我们对60只荷瘤裸鼠、5名健康志愿者和13例PCa患者的外周血和尿液样本中的SATB-1阳性CTC进行了分离、鉴定和计数。结合临床病理因素，分析了该系统的临床价值，并评估了SATB-1阳性CTC在PCa诊断中的可能性。结果表明，本研究构建的前列腺癌CTC分选与鉴定系统对PBS、尿液和血液模拟样本中CTC的回收率均超过85%。SATB-1在不同PCa细胞系中的表达水平不同，在高侵袭性的PCa DU-145细胞系中相对较高。不同临床特征的PCa患者血液样本以及部分骨转移PCa患者尿液样本中CTC的SATB-1表达不同，外周血的检测灵敏度高于尿液。本研究对于基于CTC计数和CTC中SATB-1表达的PCa早期诊断及骨转移评估具有重要的临床参考价值。
