Zhong Yuan Academy of Biological Medicine, Liaocheng People's Hospital, Liaocheng, Shandong 252000, China.
Department of thoracic Surgery, Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250000, China; Department of Thoracic Surgery, Liaocheng People's Hospital, Liaocheng, Shandong 252000, China.
Clin Chim Acta. 2018 Oct;485:95-102. doi: 10.1016/j.cca.2018.06.034. Epub 2018 Jun 22.
Circulating tumor cells (CTCs) provide an opportunity to obtain pivotal biological information required for the development of personalized medicine. However, the current assays of CTCs' detection face serious challenges regarding specificity and sensitivity.
In this study, we developed a novel strategy that combined negative enrichment (NE), immunocytochemistry CD45 staining and fluorescence in situ hybridization (FISH) to identify, enumerate and characterize CTCs. CTCs were identified as DAPI+/CD45-/Chromosome multiploid. The assay was evaluated with different cancer cell lines including lung, breast, esophageal and gastric cancer. And then, the developed assay was applied in cancer patients to explore the possibility of clinical application and whether CTC number was related to clinicopathological factors.
The average recover rate of esophageal cancer cell line Eca-109 using negative enrichment was higher than 80% and the multiploid cells rate of four cancer cell lines were >96%, which demonstrate the NE-FISH platform is favorable for CTCs detection. CTCs count was significantly higher in lung cancer patients than healthy controls and benign lung disease with an area under ROC curve of 0.905 (95% confidence interval 0.866-0.944, P < .001). Using a cutoff value of 2 CTCs, the positive rate of detecting lung, gastric, breast and esophageal cancer patients were 71.33%, 86.21%, 76.77% and 78.35%, respectively. Besides, CTCs could be detected in stage I with the positive rate of 64.15% for lung cancer, 83.33% for gastric cancer, 78.95% for breast cancer and 68.18% for esophageal cancer, which may promote the early diagnose and influence the treatment decision for better management of those cancer in clinic.
Our study showed that CTCs could be detected in diverse cancers using the novel NE-FISH platform with high sensitivity and specificity. Therefore, analysis of CTCs with NE-FISH has a clear potential to improve the management of cancer patients in clinical use.
循环肿瘤细胞（CTC）为获取个性化医学所需的关键生物学信息提供了机会。然而，目前的 CTC 检测方法在特异性和敏感性方面面临严峻挑战。
在这项研究中，我们开发了一种新的策略，将负向富集（NE）、免疫细胞化学 CD45 染色和荧光原位杂交（FISH）相结合，以识别、计数和表征 CTC。CTC 被鉴定为 DAPI+/CD45-/染色体多倍体。该检测方法在不同的癌细胞系（包括肺癌、乳腺癌、食管癌和胃癌）中进行了评估。然后，将开发的检测方法应用于癌症患者，以探索其临床应用的可能性以及 CTC 数量是否与临床病理因素有关。
食管癌细胞系 Eca-109 采用负向富集的平均回收率高于 80%，四种癌细胞系的多倍体细胞率均>96%，表明 NE-FISH 平台有利于 CTC 的检测。肺癌患者的 CTC 计数明显高于健康对照组和良性肺部疾病患者，ROC 曲线下面积为 0.905（95%置信区间 0.866-0.944，P<.001）。使用 2 个 CTC 的截断值，肺癌、胃癌、乳腺癌和食管癌患者的阳性检出率分别为 71.33%、86.21%、76.77%和 78.35%。此外，在 I 期肺癌、83.33%的胃癌、78.95%的乳腺癌和 68.18%的食管癌患者中也可以检测到 CTC，这可能有助于早期诊断，并影响治疗决策，从而更好地管理临床癌症患者。
我们的研究表明，使用新型 NE-FISH 平台可以高度敏感和特异性地检测多种癌症中的 CTC。因此，NE-FISH 分析 CTC 具有明显的潜力，可改善癌症患者的临床管理。
