血浆循环游离DNA水平及其完整性用于肺结节良恶性的诊断研究

A clinical study of circulating free DNA concentration and integrity in peripheral blood in the diagnosis of benign and malignant pulmonary nodules

  • 摘要:
    目的 探讨血浆循环游离DNA(cfDNA)水平及其完整性对肺结节良恶性的诊断价值。
    方法 选取110例肺结节患者作为研究对象,根据病理诊断结果分为良性肺结节组60例与恶性肺结节组50例,另选取同期健康体检者30例设为对照组。采用酶联免疫吸附试验检测血浆癌胚抗原(CEA)、细胞角蛋白19片段抗原21-1(CYFRA21-1)水平,采用实时荧光定量聚合酶链反应(qRT-PCR)检测血浆cfDNA水平和cfDNA完整性; 绘制受试者工作特征(ROC)曲线,分析并比较血浆cfDNA水平、cfDNA完整性、CEA、Cyfra21-1单独和联合应用对恶性肺结节的诊断效能。
    结果 恶性肺结节组血浆cfDNA、CEA、Cyfra21-1水平依次为1 154.83(452.85, 1 642.31)、7.93(3.21, 10.31)、5.75(2.85, 8.12) ng/mL, 分别高于良性肺结节组的385.43(176.45, 704.55)、2.67(1.36, 5.45)、2.74(1.43, 3.96) ng/mL, 差异有统计学意义(P < 0.001); 恶性肺结节组患者cfDNA完整性为0.68(0.47, 0.91), 高于良性肺结节组的0.37(0.26, 0.59), 差异有统计学意义(P < 0.001)。血浆cfDNA水平和cfDNA完整性与恶性肺结节患者性别、年龄、是否吸烟、肿瘤直径、病理类型、TNM分期、肿瘤分化程度、淋巴结转移均无相关性(P>0.05)。ROC曲线显示,血浆cfDNA水平、cfDNA完整性诊断恶性肺结节的曲线下面积、敏感度、特异度均大于或高于CEA、Cyfra21-1; 血浆cfDNA、CEA、Cyfra21-1水平和cfDNA完整性联合诊断恶性肺结节的敏感度、特异度均高于四者单独检测,差异有统计学意义(P < 0.05)。
    结论 血浆cfDNA水平和cfDNA完整性对恶性肺结节具有一定的诊断价值,可作为肺结节良恶性辅助诊断的分子生物学指标。

     

    Abstract:
    Objective  To explore the concentration and integrity of circulating free DNA (cfDNA) in plasma, and its diagnostic value for benign and malignant pulmonary nodules.
    Methods  According to pathological diagnosis results, the patients were divided into benign pulmonary nodules group(60 cases) and malignant pulmonary nodule group(50 cases), and another 30 healthy examiners in the same period were selected as control group. Plasma carcinoembryonic antigen (CEA) and cytokeratin fragment 19 (Cyfra21-1) expression levels were detected by enzyme-related immunosorbent assay (ELISA). The cfDNA level and cfDNA integrity were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Receiver operating characteristic (ROC) curve was drawn, and the diagnostic value of cfDNA, CEA and Cyfra21-1 in NSCLC was analyzed by ROC curve, and the diagnostic efficacy of each index and their combination was compared.
    Results  Compared with the benign pulmonary nodule group 385.43 (176.45, 704.55), 0.37 (0.26, 0.59) ng/mL, 2.67 (1.36, 5.45) ng/mL, 2.74 (1.43, 3.96) ng/mL, the plasma levels of cfDNA 1 154.83 (452.85, 1 642.31) ng/mL, cfDNA integrity0.68 (0.47, 0.91) ng/mL, CEA7.93 (3.21, 10.31) ng/mL and Cyfra21-1 5.75 (2.85, 8.12) ng/mL in the malignant pulmonary nodule group were significantly increased (P < 0.05). Plasma cfDNA concentration and integrity showed no significant correlation with gender, age, smoking or not, tumor diameter, pathological type, TNM stage, degree of tumor differentiation and lymph node metastasis in lung cancer patients (P>0.05). ROC curve showed that area under the curve, sensitivity and specificity of plasma cfDNA concentration and cfDNA integrity were higher or larger than those of CEA and Cyfra21-1, the sensitivity and specificity of plasma cfDNA concentration and cfDNA integrity combined with CEA and Cyfra21-1 in the diagnosis of NSCLC were higher than those of their single detection of plasma cfDNA concentration, cfDNA integrity, CEA, Cyfra21-1 (P < 0.05).
    Conclusion  Plasma cfDNA and its integrity have certain clinical value in the diagnosis of benign and malignant pulmonary nodules, and can be used as molecular biological indicators for the auxiliary diagnosis of benign and malignant pulmonary nodules.

     

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