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:ObjectiveTo explore the concentration and integrity of circulating free DNA (cfDNA) in plasma, and its diagnostic value for benign and malignant pulmonary nodules.
MethodsAccording 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.
ResultsCompared 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 integrity[0.68 (0.47, 0.91) ng/mL], CEA[7.93 (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).
ConclusionPlasma 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.
-
-
表 1 qRT-PCR引物序列
基因 上游引物5′-3′ 下游引物5′-3′ ALU115 CCTGAGGTCAGGAGTTCGAG CCCGAGTAGCTGGGATTACA ALU247 GTGGCTCACGCCTGTAATC CAGGCTGGAGTGCAGTGG 
下载: 导出CSV
表 2 3组血浆cfDNA水平、cfDNA完整性比较[M(P25, P75)]
组别 n cfDNA/(ng/mL) cfDNA完整性 对照组 30 265.23(80.52, 475.34)*** 0.23(0.15, 0.32)*** 良性肺结节组 60 385.43(176.45, 704.55)*** 0.37(0.26, 0.59)*** 恶性肺结节组 50 1 154.83(452.85, 1 642.31) 0.68(0.47, 0.91) 与恶性肺结节组比较, ***P < 0.001。
下载: 导出CSV
表 3 血浆cfDNA水平、cfDNA完整性与恶性肺结节患者临床病理特征的相关性[M(P25, P75)]
特征 分类 n cfDNA/(ng/mL) Z P cfDNA完整性 Z P 性别 男 30 1 054.62(405.38, 1 621.22) 1.054 0.310 0.61(0.43, 0.89) 1.198 0.274 女 20 1 108.45(434.21, 1 637.26) 0.71(0.45, 0.93) 年龄 < 50岁 28 1 035.54(396.85, 1 630.51) 2.134 0.135 0.65(0.43, 0.92) 2.025 0.183 ≥50岁 22 1 115.33(426.32, 1 658.45) 0.70(0.41, 0.90) 吸烟 是 18 1 149.55(401.21, 1 639.76) 2.987 0.072 0.67(0.39, 0.87) 1.268 0.260 否 32 1 018.53(453.66, 1 641.72) 0.63(0.42, 0.95) 肿瘤直径 < 4 cm 26 1 035.82(401.52, 1 602.42) 0.373 0.564 0.66(0.38, 0.87) 3.344 0.067 ≥4 cm 24 1 094.47(442.53, 1 640.65) 0.69(0.41, 0.95) 病理类型 腺癌 31 956.42(395.33, 1 583.22) 1.825 0.177 0.65(0.40, 0.91) 1.035 0.323 鳞癌 19 1 168.32(456.44, 1 655.78) 0.72(0.39, 0.90) TNM分期 Ⅰ~Ⅱ期 23 987.42(401.26, 1 609.58) 0.767 0.305 0.61(0.35, 0.89) 0.451 0.507 Ⅲ~Ⅳ期 27 1 165.28(441.53, 1 654.67) 0.73(0.45, 0.94) 肿瘤分化程度 低、中分化 35 1 092.41(413.46, 1 624.43) 3.153 0.069 0.70(0.44, 0.95) 0.361 0.548 高分化 15 1 123.45(398.56, 1 641.55) 0.64(0.38, 0.89) 淋巴结转移 有 29 1 110.24(473.46, 1 655.31) 1.564 0.201 0.62(0.38, 0.86) 2.856 0.091 无 21 985.36(402.21, 1 593.34) 0.74(0.47, 0.95)
下载: 导出CSV
表 4 3组血浆CEA、Cyfra21-1水平比较[M(P25, P75)]
ng/mL 组别 n CEA Cyfra21-1 对照组 30 1.78(0.52, 3.45)*** 1.69(0.22, 2.35)*** 良性肺结节组 60 2.67(1.36, 5.45)*** 2.74(1.43, 3.96)*** 恶性肺结节组 50 7.93(3.21, 10.31) 5.75(2.85, 8.12) CEA: 癌胚抗原; Cyfra21-1: 细胞角蛋白19片段抗原21-1。
与恶性肺结节组比较, ***P < 0.001。
下载: 导出CSV
表 5 血浆cfDNA、CEA、Cyfra21-1水平和cfDNA完整性对恶性肺结节的诊断效能
指标 曲线下面积 95%CI 临界值 敏感度/% 特异度/% cfDNA 0.839 0.765~0.872 687.23 ng/mL 85.92* 74.21* cfDNA完整性 0.842 0.787~0.897 0.69 81.82* 72.34* CEA 0.783 0.715~0.851 7.65 ng/mL 79.12* 70.45* Cyfra21-1 0.795 0.724~0.843 6.93 ng/mL 76.92* 71.61* 联合诊断 0.942 0.906~0.975 — 92.34 95.56 与联合诊断比较, *P < 0.05。
下载: 导出CSV
-
[1] XING W, SUN H, YAN C, et al. A prediction model based on DNA methylation biomarkers and radiological characteristics for identifying malignant from benign pulmonary nodules[J]. BMC Cancer, 2021, 21(1): 263. doi: 10.1186/s12885-021-08002-4
[2] 刘博, 黄露, 曹亚, 等. 叶酸受体阳性CTC检测在恶性肺部结节辅助诊断中的应用[J]. 第三军医大学学报, 2020, 42(23): 2330-2335. doi: 10.16016/j.1000-5404.202007074 [3] FERNANDEZ-GARCIA D, HILLS A, PAGE K R, et al. Plasma cell-free DNA (cfDNA) as a predictive and prognostic marker in patients with metastatic breast cancer[J]. Breast Cancer Res, 2019, 21(1): 149. doi: 10.1186/s13058-019-1235-8
[4] 贾勐, 李竹瑶, 秦波, 等. 血浆游离DNA完整性指标在恶性甲状腺肿瘤中的诊断价值[J]. 中华实验外科杂志, 2020, 37(7): 1294-1296. doi: 10.3760/cma.j.cn421213-20191201-01514 [5] 凌云志, 沈益青. CEA、SCC-Ag、CYFRA21-1联合检测对非小细胞肺癌的临床诊断价值[J]. 国际检验医学杂志, 2021, 42(2): 231-234. https://www.cnki.com.cn/Article/CJFDTOTAL-GWSQ202102025.htm [6] FU L, WANG R, YIN L, et al. CYFRA21-1 tests in the diagnosis of non-small cell lung cancer: A meta-analysis[J]. Int J Biol Markers, 2019, 34(3): 251-261. doi: 10.1177/1724600819868234
[7] 谢冰峰, 黎明, 朱勇军. 肺癌患者围化疗期血清CYFRA21-1、NSE和CA125水平变化的临床价值[J]. 重庆医学, 2019, 48(6): 1049-1051. doi: 10.3969/j.issn.1671-8348.2019.06.040 [8] JONNA S, SUBRAMANIAM D S. Molecular diagnostics and targeted therapies in non-small cell lung cancer (NSCLC): an update[J]. Discov Med, 2019, 27(148): 167-170.
[9] 南永刚, 王常利, 刘新芸, 等. 干细胞特征相关自身抗体对非小细胞肺癌患者早期诊断及预后评估的价值[J]. 现代肿瘤医学, 2022, 30(13): 2374-2380. doi: 10.3969/j.issn.1672-4992.2022.13.016 [10] PÖS O, BIRÓO, SZEMES T, et al. Circulating cell-free nucleic acids: characteristics and applications[J]. Eur J Hum Genet, 2018, 26(7): 937-945. doi: 10.1038/s41431-018-0132-4
[11] 李芳芳, 陶紫薇, 沈永利, 等. 基于血液DNA甲基化在肝癌诊断中的研究进展[J]. 实用临床医药杂志, 2021, 25(9): 117-122. doi: 10.7619/jcmp.20210641 [12] 王健, 董栋, 渠敬明, 等. 血浆游离DNA在非小细胞肺癌诊断中的价值分析[J]. 医药前沿, 2019, 9(34): 105-106. https://www.cnki.com.cn/Article/CJFDTOTAL-ZFBD201706011.htm [13] 王宇轩, 苏崇玉, 刘志东. Alu-qPCR检测游离DNA长度在肺结节良恶性鉴别中的应用[J]. 现代肿瘤医学, 2021, 29(6): 944-949. doi: 10.3969/j.issn.1672-4992.2021.06.008 [14] SOLIMAN S E S, ALHANAFY A M, HABIB M S E D, et al. Serum circulating cell free DNA as potential diagnostic and prognostic biomarker in non small cell lung cancer[J]. Biochem Biophys Rep, 2018, 15: 45-51.
[15] 樊庆宇, 王雅静, 仲悦娇, 等. 胃癌血浆游离DNA检测的临床意义[J]. 临床肿瘤学杂志, 2020, 25(3): 241-245. https://www.cnki.com.cn/Article/CJFDTOTAL-LCZL202003011.htm [16] 裴东方, 陈少武, 李静. 低剂量螺旋CT联合肿瘤标志物CEA, SCC-Ag, ProGRP, NSE对良恶性孤立性肺结节的鉴别诊断价值[J]. 实用癌症杂志, 2021, 36(4): 614-617. https://www.cnki.com.cn/Article/CJFDTOTAL-SYAZ202104024.htm [17] NUMATA T, ENDO T, YANAI H, et al. Serum CEA and CYFRA levels in ALK-rearranged NSCLC patients: correlation with distant metastasis[J]. In Vivo, 2020, 34(4): 2095-2100.
[18] 官燕飞, 陈健锋, 袁斌, 等. 外周血循环肿瘤细胞联合细胞角蛋白19片段21-1检测在非小细胞肺癌筛查中的应用价值[J]. 安徽医科大学学报, 2019, 54(10): 1615-1619. https://www.cnki.com.cn/Article/CJFDTOTAL-YIKE201910024.htm [19] 冯小玲, 陈红梅, 成军霞, 等. 血清NSE、CYFRA21-1、SCC水平联合CT灌注成像对肺结节良恶性病变的诊断价值[J]. 标记免疫分析与临床, 2020, 27(9): 1551-1555, 1619. https://www.cnki.com.cn/Article/CJFDTOTAL-BJMY202009022.htm [20] 范伟, 孙振宇, 孟祥宽, 等. 血清CEA、CYFRA21-1、VEGF、STK1及SF联合检测在非小细胞肺癌患者诊断中的价值分析[J]. 标记免疫分析与临床, 2020, 27(3): 480-483, 507. https://www.cnki.com.cn/Article/CJFDTOTAL-BJMY202003027.htm -
期刊类型引用(6)
1. 许洁, 周碧瑶, 李萍. 重症监护室心力衰竭患者的临床护理中应用标准化护理程序的效果观察. 中国医药科学. 2020(03): 150-152 . 
百度学术
2. 林岚. 系统呼吸护理对呼吸衰竭机械通气患者的应用价值分析. 中国医药指南. 2019(01): 246-247 . 百度学术
3. 柴华. 全面护理在重症肺炎合并急性心力衰竭中的应用. 中国继续医学教育. 2019(10): 165-167 . 百度学术
4. 张琼. 呼吸系统功能评分机械通气治疗呼吸衰竭撤机. 中国继续医学教育. 2019(14): 121-123 . 百度学术
5. 甘霞, 许静. 6个月以下婴儿肺炎病因及并发心力衰竭的治疗探讨. 中外医疗. 2018(20): 25-27+75 . 百度学术
6. 张稳. 优化个体护理对ICU重症肺炎患者生理指标改善的影响. 实用临床护理学电子杂志. 2018(07): 30+33 . 百度学术
其他类型引用(1)
计量
- 文章访问数: 185
- HTML全文浏览量: 58
- PDF下载量: 13
- 被引次数: 7
苏公网安备 32100302010246号
