Wintachai Preawwalee, Lim Jing Quan, Techasen Anchalee, Lert-Itthiporn Worachart, Kongpetch Sarinya, Loilome Watcharin, Chindaprasirt Jarin, Titapun Attapol, Namwat Nisana, Khuntikeo Narong, Jusakul Apinya
Biomedical Sciences Program, Graduate School, Khon Kaen University, Khon Kaen 40002, Thailand.
Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen 40002, Thailand.
Diagnostics (Basel). 2021 May 30;11(6):999. doi: 10.3390/diagnostics11060999.
The analysis of cfDNA has been applied as a liquid biopsy in several malignancies. However, its value in the diagnosis and prognosis of cholangiocarcinoma (CCA) have not been well defined. We aimed to investigate the diagnostic and prognostic values of cfDNA level and tumor-specific mutation in circulating DNA (ctDNA) in CCA. The plasma cfDNA levels from 62 CCA patients, 33 benign biliary disease (BBD) patients and 30 normal controls were quantified by fluorescent assay. Targeted probe-based sequencing of 60 genes was applied for mutation profiling in 10 ctDNA samples and their corresponding treatment-naïve tissues. cfDNA levels in CCA were significantly higher than those in BBD and normal controls. We found that cfDNA levels at 0.2175 and 0.3388 ng/µL significantly discriminated CCA from healthy controls and BBD with 88.7 and 82.3% sensitivity and 96.7 and 57.6% specificity, respectively. cfDNA levels showed superior diagnostic efficacy in detecting CCA compared to CEA and CA19-9. (30%), (30%), (30%), and (30%) mutations were the most common. Using nine frequently mutated genes in the ctDNA samples, the diagnostic accuracy of cfDNA sequencing was 90.8%, with 96.7% average sensitivity and 72.4% specificity. This study supports the use of cfDNA as a diagnosis and prognostic biomarker for CCA.
循环游离DNA（cfDNA）分析已被用作多种恶性肿瘤的液体活检。然而，其在胆管癌（CCA）诊断和预后方面的价值尚未明确界定。我们旨在研究CCA患者循环DNA（ctDNA）中cfDNA水平和肿瘤特异性突变的诊断及预后价值。采用荧光分析法对62例CCA患者、33例良性胆道疾病（BBD）患者和30例正常对照者的血浆cfDNA水平进行定量分析。对10份ctDNA样本及其相应的未经治疗的组织进行基于靶向探针的60个基因测序，以进行突变谱分析。CCA患者的cfDNA水平显著高于BBD患者和正常对照者。我们发现，cfDNA水平分别为0.2175和0.3388 ng/µL时，可分别以88.7%和82.3%的灵敏度以及96.7%和57.6%的特异性显著区分CCA与健康对照者和BBD患者。与癌胚抗原（CEA）和糖类抗原19-9（CA19-9）相比，cfDNA水平在检测CCA方面显示出更高的诊断效能。（30%）、（30%）、（30%）和（30%）的突变最为常见。利用ctDNA样本中9个频繁突变的基因，cfDNA测序的诊断准确率为90.8%，平均灵敏度为96.7%，特异性为72.4%。本研究支持将cfDNA用作CCA的诊断和预后生物标志物。
