Yang Jian-Yu, Sun Yong-Wei, Liu De-Jun, Zhang Jun-Feng, Li Jiao, Hua Rong
Department of Biliary-Pancreatic Surgery, Ren Ji Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine Shanghai, 200127, China.
Am J Cancer Res. 2014 Nov 19;4(6):663-73. eCollection 2014.
Pancreatic ductal adenocarcinoma (PDAC) accounts for approximately 90-95% exocrine malignant tumors of the pancreas. The high prevalence of metastasis and the difficulty of early diagnosis lead to a dismal prognosis. MicroRNAs (miRNAs) play a critical role in extensive biological processes. The purpose of this study was to evaluate the feasibility of stool miRNAs as novel biomarker for PDAC screening. MiRNAs were extracted from clinical specimens which included cancer and matched adjacent benign pancreatic tissues of 30 PDAC patients, pancreatic juice of 20 from the 30 PDAC patients and 10 chronic pancreatitis (CP) patients, stool samples of the 30 PDAC patients, the 10 CP patients and 15 healthy volunteers. Relative expression of a panel of 5 dysregulated miRNAs (miR-21, miR-155, miR-196a, miR-216 and miR-217) was analyzed with qRT-PCR. Receiver operating characteristic curve (ROC) analysis was performed to assess the diagnosing value of stool miRNAs in PDAC patients. The study showed that our methods of extracting and detecting miRNAs from pancreatic juice and stool specimens had high reproducibility. Compared to matched adjacent benign pancreatic tissues and pancreatic juice of CP patients, the expression of miR-21 (P = 0.0021 and P = 0.0027) as well as miR-155 (P = 0.0087 and P = 0.0067) was significantly higher and the expression of miR-216 (P < 0.0001 and P = 0.0044) was significantly lower in primary tumor tissues and pancreatic juice of PDAC patients. PDAC patients had a significantly higher stool miR-21 and miR-155 (P = 0.0049 and P = 0.0112) and lower miR-216 level (P = 0.0002) compared to normal controls. The same results were obtained in the expression levels of stool miR-21, miR-155 and miR-216 between PDAC and CP patients (P = 0.0337, P = 0.0388 and P = 0.0117, respectively). Receiver operating characteristic (ROC) analysis by using stool miRNAs expression indicated that combination of miR-21 and miR-155 had best sensitivity of 93.33% while the combination of miR-21, miR-155 and miR-216 would be best for detecting and screening PDAC with area under the curve (AUC) of 0.8667 (95% CI: 0.7722-0.9612) and a better balance of sensitivity and specificity (83.33% vs. 83.33%). Our data indicate that miRNAs could be extracted and detected from pancreatic juice and stool efficiently and reproducibly. MiR-21, miR-155 and miR-216 in stool have the potential of becoming biomarkers for screening PDAC.
胰腺导管腺癌（PDAC）约占胰腺外分泌恶性肿瘤的90 - 95%。转移的高发生率和早期诊断的困难导致预后不佳。微小RNA（miRNA）在广泛的生物学过程中起关键作用。本研究的目的是评估粪便miRNA作为PDAC筛查新生物标志物的可行性。从临床标本中提取miRNA，这些标本包括30例PDAC患者的癌组织及配对的相邻胰腺良性组织、30例PDAC患者中的20例以及10例慢性胰腺炎（CP）患者的胰液、30例PDAC患者、10例CP患者和15名健康志愿者的粪便样本。用qRT-PCR分析一组5种失调miRNA（miR - 21、miR - 155、miR - 196a、miR - 216和miR - 217）的相对表达。进行受试者操作特征曲线（ROC）分析以评估粪便miRNA对PDAC患者的诊断价值。研究表明，我们从胰液和粪便标本中提取和检测miRNA的方法具有高重现性。与配对的相邻胰腺良性组织及CP患者的胰液相比，PDAC患者的原发肿瘤组织和胰液中miR - 21（P = 0.0021和P = 0.0027）以及miR - 155（P = 0.0087和P = 0.0067）的表达显著更高，而miR - 216（P < 0.0001和P = 0.0044）的表达显著更低。与正常对照相比，PDAC患者粪便中miR - 21和miR - 155显著更高（P = 0.0049和P = 0.0112），miR - 216水平更低（P = 0.0002）。PDAC患者与CP患者粪便中miR - 21、miR - 155和miR - 216的表达水平也得到相同结果（分别为P = 0.0337、P = 0.0388和P = 0.0117）。使用粪便miRNA表达进行的受试者操作特征（ROC）分析表明，miR - 21和miR - 155组合的敏感性最佳，为93.33%，而miR - 21、miR - 155和miR - 216组合对检测和筛查PDAC最佳，曲线下面积（AUC）为0.8667（95% CI：0.7722 - 0.9612），敏感性和特异性平衡更好（83.33%对83.33%）。我们的数据表明，可以从胰液和粪便中高效且可重复地提取和检测miRNA。粪便中的miR - 21、miR - 155和miR - 216有成为筛查PDAC生物标志物的潜力。
