CAJ | 학술논문

目的探讨原发性胆汁性肝硬化(PBC)患者血浆微小RNA(miRNA)的表达及临床意义.方法采用随机对照研究的方法,收集上海松江中心医院2010年12月至2013年1月就诊的19例PBC患者的血浆,同时收集了性别年龄与PBC患者相匹配的10名健康体检者和10例病毒性肝炎患者的血浆,从中选取3例PBC患者和3名健康体检者,应用芯片技术进行miRNA表达谱筛选.针对筛选出有表达差异的miRNA,采用荧光定量PCR(qRT-PCR)技术对收集的3组标本进行验证,统计学分析和绘制受试者工作曲线(ROC)评估差异miRNA的临床价值.结果芯片结果显示PBC组与对照组共有16个差异表达miRNA.qRT-PCR验证发现,与正常组相比,PBC组miRNA-572和miRNA-575表达明显上调(P＜0.05),miRNA-92a-3p和miRNA-4516表达明显下调(P＜0.05).而与非PBC肝硬化组相比,PBC组miRNA-92a-3p和miRNA-4516表达下调(P＜0.05).miRNA-92a-3p在评价PBC组和健康对照组的曲线下面积(AUC)为0.92(0.86～ 1.00),当最佳临界值为1.26,敏感度92％,特异度80％.miRNA-4516在评价PBC组和健康对照组的曲线下面积为0.89(0.75 ～1.00),当最佳临界值为1.16,敏感度85％,特异度70％.miRNA-92a-3p和miRNA-4516在鉴别诊断PBC和非PBC肝硬化时曲线下面积分别为0.84 (0.68～0.99)和0.76(0.57 ～0.96).当miRNA-92a-3p取最佳临界值为1.08,敏感度89％,特异度81％.当miRNA-4516取最佳临界值为1.06,敏感度77％,特异度68％.结论 PBC患者血浆中特异表达的miRNA对PBC诊断及鉴别诊断的具有潜在临床应用价值.
목적 탐토원발성담즙성간경화(PBC)환자혈장미소RNA(miRNA)적표체급림상의의.방법 채용수궤대조연구적방법,수집상해송강중심의원2010년12월지2013년1월취진적19례PBC환자적혈장,동시수집료성별년령여PBC환자상필배적10명건강체검자화10례병독성간염환자적혈장,종중선취3례PBC환자화3명건강체검자,응용심편기술진행miRNA표체보사선.침대사선출유표체차이적miRNA,채용형광정량PCR(qRT-PCR)기술대수집적3조표본진행험증,통계학분석화회제수시자공작곡선(ROC)평고차이miRNA적림상개치.결과 심편결과현시PBC조여대조조공유16개차이표체miRNA.qRT-PCR험증발현,여정상조상비,PBC조miRNA-572화miRNA-575표체명현상조(P＜0.05),miRNA-92a-3p화miRNA-4516표체명현하조(P＜0.05).이여비PBC간경화조상비,PBC조miRNA-92a-3p화miRNA-4516표체하조(P＜0.05).miRNA-92a-3p재평개PBC조화건강대조조적곡선하면적(AUC)위0.92(0.86～ 1.00),당최가림계치위1.26,민감도92％,특이도80％.miRNA-4516재평개PBC조화건강대조조적곡선하면적위0.89(0.75 ～1.00),당최가림계치위1.16,민감도85％,특이도70％.miRNA-92a-3p화miRNA-4516재감별진단PBC화비PBC간경화시곡선하면적분별위0.84 (0.68～0.99)화0.76(0.57 ～0.96).당miRNA-92a-3p취최가림계치위1.08,민감도89％,특이도81％.당miRNA-4516취최가림계치위1.06,민감도77％,특이도68％.결론 PBC환자혈장중특이표체적miRNA대PBC진단급감별진단적구유잠재림상응용개치.
Objective To evaluate the expression and clinical significance of miRNA in plasma of patients with primary biliary cirrhosis.Methods Plasma from 19 PBC patients,10 healthy volunteers and 10 viral hepatitis patients were selected from Shanghai Songjiang Hospital from december 2010 to January 2013.Among them 3 PBC patients' plasma and 3 healthy volunteers' plasma were detected by miRNA microarray for miRNA expression profile examination.Real-time PCR was used to verify the results of microarray,miRNA target gene predictior software was used to predict the target genes of differentially expressed miRNA.ROC was used to determine the clinical value of plasma miRNA.Results According to microarray,a total of 16 miRNAs were found to be differentially expressed.As revealed by qRT-PCR,the expression of miRNA-92a-3p and miRNA-4516 decreased while the expression of miRNA-572 and miRNA-575 were up-regulated in PBC group compared with healthy controls (P ＜ 0.05).In comparison with nonPBC cirrhosis group,only miRNA-92a-3p and miRNA-4516 were down-regulated (P ＜ 0.05).The area under the curve (AUC)of miRNA-92a-3p for the diagnosis and differential diagnosis of PBC were 0.92 and 0.84,respectively.while for The area under the ROC curve of miRNA-4516,the AUC for diagnosis and differential diagnosis PBC were 0.89 and 0.76,respectively.The optimal cut-off values for identifying PBC from healthy controls were defined as 1.26 ng/μl.for miRNA-92a-3p (sensitivity,92％ ;specificity,80％)and 1.16ng/ul for miRNA-4516 (sensitivity,85％ ;specificity,70％)respectively.The optimal cut-off values for identifying PBC from viral hepatitis were defined as 1.08 ng/μl.for miRNA-92a-3p (sensitivity,89％ ; specificity,81％)and 1.06 ng/μl for miRNA-4516 (sensitivity,77％ ;specificity,68％).Conclusion The results indicate that plasma from patients with PBC has a unique miRNA exprssion profile and these differentially expressed miRNA can be used as clinical biomarkers of PBC.