Kolostova Katarina, Pinkas Michael, Jakabova Anna, Pospisilova Eliska, Svobodova Pavla, Spicka Jan, Cegan Martin, Matkowski Rafal, Bobek Vladimir
Department of Laboratory Genetics, University Hospital Kralovske Vinohrady Srobarova 50, 10034 Prague, Czech Republic.
Department of Pathology and Thoracic Surgery, Masaryk's Hospital in Usti nad Labem Krajska zdravotni a.s., Socialni pece 3316/12A, 40113 Usti nad Labem, Czech Republic.
Am J Cancer Res. 2016 May 1;6(5):973-80. eCollection 2016.
The main focus of the study was to detect circulating tumor cells (CTCs) in ovarian cancer (OC) patients using a new methodological approach (MetaCell(TM)) which is based on size-dependent separation of CTCs and subsequent cytomorphological evaluation. Cytomorphological evaluation using vital fluorescence microscopy approach enables to use the captured cells for further RNA/DNA analysis. The cytomorphological analysis is then completed by gene expression analysis (GEA). GEA showed that relative expression of EPCAM is elevated in CTC-enriched fractions in comparison to the whole peripheral blood sample and that the expression grows with in vitro cultivation time. Comparison of the relative gene expression level in the group of peripheral blood samples and CTC-fraction samples confirmed a statistically significant difference for the following genes (p < 0.02): KRT7, WT1, EPCAM, MUC16, MUC1, KRT18 and KRT19. Thus, we suggest that the combination of the above listed genes could confirm CTCs presence in OC patients with higher specificity than when GEA tests are performed for one marker only. The GEA revealed two separate clusters identifying patients with or without CTCs.
该研究的主要重点是使用一种新的方法(MetaCell™)检测卵巢癌(OC)患者循环肿瘤细胞(CTC),该方法基于CTC的大小依赖性分离及随后的细胞形态学评估。采用活细胞荧光显微镜方法进行细胞形态学评估能够将捕获的细胞用于进一步的RNA/DNA分析。然后通过基因表达分析(GEA)完成细胞形态学分析。GEA显示,与全外周血样本相比,EPCAM在富含CTC的组分中的相对表达升高,且该表达随体外培养时间增加。外周血样本组和CTC组分样本组的相对基因表达水平比较证实,以下基因存在统计学显著差异(p < 0.02):KRT7、WT1、EPCAM、MUC16、MUC1、KRT18和KRT19。因此,我们认为,与仅对一种标志物进行GEA检测相比,上述基因组合能够以更高的特异性确认OC患者中CTC的存在。GEA揭示了两个独立的聚类,分别识别有或无CTC的患者。