粪便microRNAs检测用于胰腺癌筛查诊断的价值评价
Expression of microRNAs in fecal of patients for pancreatic cancer screening and detection
摘要目的 检测胰腺癌患者粪便microRNAs,评价其诊断价值.方法 收集29例胰腺癌患者、22例慢性胰腺炎(CP)患者以及13例健康志愿者的粪便标本,抽提粪便总RNA,应用实时定量PCR法检测各组样本miR-21、miR-155、miR-181a、miR-181b、miR-196a、miR-210的表达量,以miR-16作为内参基因.应用接受者操作特征(ROC)曲线(AUC)评估microRNAs对胰腺癌的诊断价值.结果 粪便总RNA抽提及microRNAs检测方法具有稳定及可重复性.胰腺癌组miR-181b、miR-196a、miR-210的表达量分别为2.22±0.64、2.78±0.14、5.55±0.38;CP组为1.42±0.39、3.88±0.85、5.39±0.69;对照组为0.32±0.40、1.14±0.98、4.23±0.99.胰腺癌组和CP组均较对照组显著增加(P值均<0.05);而胰腺癌组与CP组间无显著差异.胰腺癌组对对照组的miR-181b AUC为0.745(95%CI 0.597~0.894),诊断胰腺癌的敏感性和特异性分别为84.6%和51.7%;miR-210的AUC为0.772(95%CI 0.629~0.914),对胰腺癌的诊断敏感性和特异性分别为84.6%和65.5%.两组比较差异均有统计学意义(P值均<0.05).miR-196a对胰腺癌无诊断意义,但胰腺癌患者粪便miR-196a的表达与肿瘤直径相关(r=0.516,P=0.041).结论 粪便RNA的抽提和microRNAs检测为无创性,且具有可重复性.miR-181b和miR-210在胰腺癌患者粪便中的表达增高,有可能是胰腺癌潜在的分子标志物.
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abstractsObjective To detect the microRNAs in fecal with patients of pancreatic cancer, and evaluate its diagnostic value. Methods Stool samples were collected from three group persons including 29 pancreatic cancer, 22 chronic pancreatitis and 13 normal controls. The total fecal microRNAs were extracted.The quantity of miR-16, miR-21, miR-155, miR-181a, miR-181b, miR-196a, and miR-210 were detected by using real-time PCR, and miR-16 was used as reference gene. ROC AUC was used to evaluate the diagnostic value for pancreatic cancer. Results MicroRNAs were efficiently obtained from stools, and independent experiments showed high reproducibility for microRNAs extraction and detection. The expression of miR-181b,miR-196a, miR-210 in fecal was 2.22 ±0.64,2.78 ±0.14, 5.55 ±0.38 in pancreatic cancer; 1.42 ±0.39,3.88 ± 0.85,5.39 ± 0.69 in chronic pancreatitis; 0.32 ± 0.40, 1.14 ± 0.98,4.23 ± 0. 99 in normal controls;the three microRNA expressions in pancreatic cancer were group and CP group significantly higher than those in normal controls ( P < 0.05 ). But there was no significant difference between pancreatic cancer group and chronic pancreatitis group. AUC of pancreatic cancer / normal controls miR 18lb was 0.745(95% CI 0. 597-0.894), the sentivity, specificity for pancreatic cancer was 84.6% and 51.7%. AUC of miR-210 was 0. 772(95% CI0.629-0.914), the sentivity, specificity for pancreatic cancer was 84.6% and 65.5%, and the difference was statistically significant (P <0.05). miR-196a was no significant for the diagnosis of pancreatic cancer, but the expression of miR-196a was correlated with the tumor size (r = 0.516, P = 0.041 ).Conclusions The extraction and detection of the fecal microRNAs were non-invasive and reproducible. The expression of miR-181b and miR-210 was increased in stool of patients with pancreatic cancer, and may be potential biomarker for pancreatic cancer.
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