Fidelia-Lambert M N, Zhuang Z, Tsokos M
Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Hum Pathol. 1999 Jan;30(1):78-80. doi: 10.1016/s0046-8177(99)90304-0.
Disseminated disease is very important in the clinical assessment of pediatric sarcomas. Several reports suggest that reverse transcriptase polymerase chain reaction (RT-PCR) holds great promise in the early staging of cancer patients in general. However, the complexities of these protocols hamper adequate standardization, and their application as routine diagnostic tools has been difficult. The aim of this study is to assess the actual minimal number of tumor cells that may be detected by RT-PCR in a blood sample. Specific tumor cell dilutions from a Ewing's sarcoma cell line reconstituted in peripheral blood from healthy individuals were "ficolled" and submitted to RNA extraction for cDNA preparation and PCR amplification of the t(11-22) (q24;q12) fusion transcript. After PCR amplification, we were able to detect the EWS/FI-1 chimeric gene product at a dilution of 10 tumor cells per 1 or 2 mL of blood. Our simple method supports a role for routine clinical use of RT-PCR in the detection of circulating Ewing's sarcoma cells.
播散性疾病在小儿肉瘤的临床评估中非常重要。几份报告表明，逆转录酶聚合酶链反应（RT-PCR）总体上在癌症患者的早期分期中具有很大前景。然而，这些方案的复杂性阻碍了充分的标准化，并且将其用作常规诊断工具一直很困难。本研究的目的是评估血液样本中通过RT-PCR可能检测到的实际肿瘤细胞最小数量。从健康个体外周血中重构的尤因肉瘤细胞系中进行特定肿瘤细胞稀释，经“ficoll”处理后进行RNA提取以制备cDNA，并对t(11-22) (q24;q12)融合转录本进行PCR扩增。PCR扩增后，我们能够在每1或2 mL血液中10个肿瘤细胞的稀释度下检测到EWS/FI-1嵌合基因产物。我们的简单方法支持RT-PCR在常规临床中用于检测循环中的尤因肉瘤细胞。
