血清循环微小RNA(c-miRNA)是癌症诊断的有用生物标志物。这里,我们描述了一步分支滚环扩增(BRCA)方法的发展,以测量血清c-miRNAs水平,以早期诊断乳腺癌。四个c-miRNAs,c-miRNA16(c-miR-16),c-miRNA21(c-miR-21),c-miRNA155(c-miR-155),从49例乳腺癌患者和19例健康对照者血清中分离出c-miRNA195(c-miR-195)。其中,45例乳腺癌患者和15例健康对照者使用一步法BRCA进行分析,4名乳腺癌患者和4名健康对照通过定量实时PCR分析[校正]。I期乳腺癌患者血清c-miR16、c-miR21、c-miR155和c-miR195水平高于健康对照组(P<0.0001)。这些水平在几种乳腺癌分子亚型中也较高(HER-2过表达,管腔A,管腔B,和三阴性乳腺癌)比健康对照受试者。c-miR16,c-miR21,c-miR155和c-miR195对乳腺癌早期诊断的诊断准确性已通过受试者工作特征(ROC)曲线测定证实。这些结果表明,BRCA方法可用于测量血清c-miRNAs水平,并且该方法具有很高的准确性,灵敏度,和特异性。此外,BRCA方法和实时定量PCR(qRT-PCR)方法均表明血清c-miR16、c-miR21、c-miR155和c-miR195水平可作为提高乳腺癌早期诊断水平的生物标志物,区分不同的乳腺癌分子亚型。
Serum circulating microRNAs (c-miRNAs) are serving as useful biomarkers for cancer diagnosis. Here, we describe the development of a one-step branched rolling circle amplification (BRCA) method to measure serum c-miRNAs levels for early diagnosis of breast cancer.
Four c-miRNAs, c-miRNA16 (c-miR-16), c-miRNA21 (c-miR-21), c-miRNA155 (c-miR-155), and c-miRNA195 (c-miR-195) were isolated from
the serum of 49 breast cancer patients and 19 healthy controls. Among them, 45 breast cancer patients and 15 healthy controls were analyzed
using one-step BRCA, 4 breast cancer patients and 4 healthy controls were analyzed by quantitative real-time PCR assay [corrected].
The serum levels of c-miR16, c-miR21, c-miR155, and c-miR195 were higher (P < 0.0001) in stage I breast cancer patients than healthy controls. These levels were also higher in several breast cancer molecular subtypes (HER-2 over-expression, Luminal A, Luminal B, and triple negative breast cancer) than in healthy control subjects. The diagnostic accuracy of c-miR16, c-miR21, c-miR155, and c-miR195 for early diagnosis of breast cancer was confirmed by receiver operating characteristic (ROC) curve assay. These results show that the BRCA method can be used to measure serum c-miRNAs levels, and that this method has high accuracy, sensitivity, and specificity. Moreover, both BRCA approach and quantitative real-time PCR (qRT-PCR) method show that the serum levels of c-miR16, c-miR21, c-miR155, and c-miR195 could be used as biomarkers to improve the early diagnosis of breast cancer, and distinguish different breast cancer molecular subtypes.