摘要
目的 miRNA是一类约高度保守的小分子非编码RNA,具有调控细胞分化、增殖、凋亡的功能,与心血管疾病密切相关。因肺动脉高压是血管增殖性疾病,与血管再生相关的miR-19a可能与肺动脉高压密切相关,循环miRNA可作为先天性心脏病相关肺动脉高压的无创诊断标志物。方法收集室间隔缺损合并重度肺动脉高压患者血浆14例(病例组),室间隔缺损不伴肺动脉高压患者血浆14例(对照组)。分别从血浆中提取总RNA并进行质量检测,进行逆转录反应,获得相应miRNA的cDNA,以U6snRNA为内参进行标定,进行实时荧光定量PCR反应,绘制标准曲线,根据标准曲线和各样本的定量PCR结果计算miR-19a的表达量并绘制ROC曲线。结果两组样本的总RNA均有较好的质量,可以满足miRNA芯片检测和实时定量PCR的实验要求。病例组血浆中miR-19a拷贝数明显高于对照组。ROC曲线下面积为0.865(95%可信区间,CI=0.727-1.002,P<0.001),循环miR-19a诊断先天性心脏病相关肺动脉高压具有较高的敏感性和特异性。结论先天性心脏病相关肺动脉高压患者血浆中miR-19a显著增高,循环miR-19a可作为先天性心脏病相关肺动脉高压新的无创性诊断标志物。
Objective To Objective MicroRNAs are small noncoding transcripts involved in many cellular mechanisms,including cardiovascular diseases.MiR-19a,in particular,was considered play a great role in antiangiogensis.Because pulmonary arterial hypertension(PAH) is an angioproliferative disease,we hypothesized that miR-19a may serve as a diagnostic marker for screening for PAH.Methods A cohort of 14 severe PAH patients and 14 age-matched controls was used in this study.Plasma samples were obtained from newly diagnosed ventricular septal defect with severe PAH and age-matched non-PAH controls.MiRNA was extracted directly from plasma and reverse-transcribed to complementary DNA.A known quantity of synthetic U6 was added for normalization.MiR-19a was then measured using quantitative reverse transcription polymerase chain reaction.The same isolation and assay were performed using a series of concentrations of U6 to make standard curve.Receiver-operator characteristic curve(ROC) analysis was used to evaluate the diagnostic ability of miR-19a for PAH.Results MiR-19a was reliably detected and quantif ied,with a statistically signif icant increase in expression in severe PAH patients compared with normal controls.The area under ROC curve(AUC) was 0.865 for separation between controls and PAH patients(95% CI,0.727 ~ 1.002,P0.001).Conclusions In summary,circulating miR-19a levels are elevated in PAH patients and may potentially serve as a useful biomarker for PAH.
出处
《中国分子心脏病学杂志》
CAS
2010年第5期285-288,共4页
Molecular Cardiology of China
基金
国家"十一五"科技支撑计划项目(2006BA101A08)
关键词
循环miRNA
肺动脉高压
先天性心脏病
Circulating miRNA
Pulmonary arterial hypertension
Congenital heart disease