纳米磁分离实时荧光RT-PCR检测食管鳞状细胞癌患者血清miR-21表达的价值?

目的：建立食管鳞状细胞癌( ESCC)患者血清中miR-21纳米磁分离( MS)实时荧光RT-PCR检测方法,观察ESCC患者血清miR-21的表达。方法分别采用Trizol法和MS法提取60例ESCC患者及60例健康体检者血清总RNA,采用实时荧光RT-PCR检测miR-21的表达。利用Bland-Altman法检验Trizol法与MS法提取的血清中miR-21表达量的一致性,利用ROC曲线评价MS法提取的miR-21表达量筛查ESCC的效能。结果 MS法能够提取血清中miR-21,其灵敏度与Trizol方法相当,Bland-Altman法检验结果显示两种方法提取的miR-21表达量具有很好的一致性。 ESCC患者miR-21表达量明显高于健康体检者( P<0．05)。 ROC曲线下面积为0．878,最佳临界值为1．115,此时敏感度为85．00%,特异度为81．67%。 ESCC患者血清miR-21的表达与TNM分期以及淋巴结转移情况相关( P<0．05)。结论 MS实时荧光RT-PCR在血清miRNA检测中有着很大的应用价值,血清miR-21的表达水平有可能作为食管癌诊断一个新的分子生物学指标。

Values of magnetic nanoparticles separation-based real-time fluorescent RT-PCR assay for detection of serum ;miR-21 in patients with esophageal squamous cell carcinoma

Objective To establish the magnetic nanoparticles separation ( MS) -based real-time fluorescent RT-PCR assay for detection of serum micR-21 in patients with esophageal squamous cell carcinoma ( ESCC) , and to reveal the expression of miR-21 in serum of patients with ESCC. Methods Sixty cases of ESCC and 60 cases of healthy individuals were collected, and total serum RNA was extracted by Trizol and MS, respectively. The expression of miR-21 was detected by real-time fluorescent RT-PCR. The consistency of miR-21 expression between Trizol and MS ex-traction was detected by Bland-Altman method. Utility of miR-21 expression detected by MS in screening ESCC was e-valuated by ROC curves. Results The MS-based real-time RT-PCR was effective for separating and extracting serum miR-21, with sensitivity similar to the Trizol-based real-time RT-PCR. Bland-Altman results showed that the ex-pression of miR-21 extraced by the two methods had good consistency. The expression of miR-21 in the serum of ESCC patients was significantly higher than that in healthy individuals (P<0. 05). The area under the ROC curve was 0. 878. 1. 115 was the best critical value, where the sensitivity and specificity were 85. 00% and 81. 67%, respectively. The ex-pression of miR-21 in ESCC patients was correlated with TNM staging and lymph node metastasis. Conclusion MS-based real-time fluorescent RT-PCR assay is of great clinical significance in detection of serum miRNA. The expression level of miR-21 in serum should be a new biomarker for the diagnosis of esophageal carcinoma.