Department of Internal Medicine 1, University of Bonn, Bonn, Germany.
PLoS One. 2012;7(8):e42141. doi: 10.1371/journal.pone.0042141. Epub 2012 Aug 7.
It is a challenging task to distinguish between benign and malignant lesions in patients with biliary strictures. Here we analyze whether determination of target gene mRNA levels in intraductal brush cytology specimens may be used to improve the diagnosis of bile duct carcinoma.
Brush cytology specimens from 119 patients with biliary strictures (malignant: n = 72; benign: n = 47) were analyzed in a retrospective cohort study. mRNA of IGF-II mRNA-binding protein 3 (IGF2BP3), homeobox B7 (HOXB7), Forkhead box M1 (FOXM1), kinesin family member 2C (KIF2C) and serine/threonine kinase NEK2 was determined by semi-quantitative RT-PCR using the ΔCt method.
IGF2BP3 (p<0.0001), HOXB7 (p<0.0001), and NEK2 (p<0.0001) mRNA expression levels were significantly increased in patients with cholangiocarcinoma or pancreatic cancer. Median ΔCt values differed by 3.5 cycles (IGF2BP3), 2.8 cycles (HOXB7) and 1.3 cycles (NEK2) corresponding to 11-fold, 7-fold and 2.5-fold increased mRNA levels in malignant versus benign samples. Sensitivity to detect biliary cancer was 76.4% for IGF2BP3 (80.9% specificity); 72.2% for HOXB7 (78.7% specificity) and 65.3% for NEK2 (72.3% specificity), whereas routine cytology reached only 43.1% sensitivity (85.4% specificity). Diagnostic precision was further improved, when all three molecular markers were assessed in combination (77.8% sensitivity, 87.2% specificity) and achieved 87.5% sensitivity and 87.2% specificity when molecular markers were combined with routine cytology.
Our data suggest that measuring IGF2BP3, HOXB7 and NEK2 mRNA levels by RT-PCR in addition to cytology has the potential to improve detection of malignant biliary disorders from brush cytology specimens.
在胆管狭窄患者中，区分良性和恶性病变是一项具有挑战性的任务。在这里，我们分析了在胆管刷取细胞学标本中测定靶基因 mRNA 水平是否可用于提高胆管癌的诊断。
在一项回顾性队列研究中，分析了 119 例胆管狭窄患者（恶性：n = 72；良性：n = 47）的刷取细胞学标本。采用半定量 RT-PCR 法，采用 ΔCt 法测定胰岛素样生长因子 II mRNA 结合蛋白 3（IGF2BP3）、同源盒 B7（HOXB7）、叉头框 M1（FOXM1）、驱动蛋白家族成员 2C（KIF2C）和丝氨酸/苏氨酸激酶 NEK2 的 mRNA。
在胆管癌或胰腺癌患者中，IGF2BP3（p<0.0001）、HOXB7（p<0.0001）和 NEK2（p<0.0001）mRNA 表达水平显著升高。恶性与良性样本相比，中位ΔCt 值相差 3.5 个循环（IGF2BP3）、2.8 个循环（HOXB7）和 1.3 个循环（NEK2），分别对应于 mRNA 水平增加 11 倍、7 倍和 2.5 倍。IGF2BP3 检测胆管癌的敏感性为 76.4%（特异性 80.9%）；HOXB7 为 72.2%（特异性 78.7%）；NEK2 为 65.3%（特异性 72.3%），而常规细胞学仅达到 43.1%的敏感性（特异性 85.4%）。当评估三种分子标志物的组合时，诊断精度进一步提高（敏感性 77.8%，特异性 87.2%），当分子标志物与常规细胞学结合时，敏感性达到 87.5%，特异性达到 87.2%。
我们的数据表明，在胆管刷取细胞学标本中，除细胞学检查外，通过 RT-PCR 测定 IGF2BP3、HOXB7 和 NEK2 mRNA 水平具有提高恶性胆道疾病检测的潜力。
