SYBR GreenⅠ荧光定量PCR检测宫颈癌血清中miR-21的表达  

Detection of expression of miR-21 in serum of cervical cancer patients by SYBR greenⅠ fluorescence quantitative PCR

作　　者：陈文璟[1] 温旺荣[1] 

机构地区：[1]暨南大学附属第一医院临床检验中心,广州510630

出　　处：《临床检验杂志》2011年第7期523-525,共3页Chinese Journal of Clinical Laboratory Science

摘　　要：目的建立SYBR GreenⅠ实时荧光定量PCR法检测宫颈疾病患者血清中miR-21的表达水平,探讨miR-21在宫颈癌临床监测中的意义。方法设计miR-21、U6茎环逆转录引物和PCR扩增引物,以U6为内参,用实时荧光定量PCR检测32例不同宫颈疾病患者血清中miR-21表达水平。结果建立的实时荧光定量PCR法能特异地检测血清中的miR-21扩增信号,熔解曲线单一,产物特异。宫颈癌患者血清中miR-21水平明显高于子宫肌瘤、宫颈炎等其他良性宫颈疾病(P<0.05),而宫颈癌患者手术切除后血清中miR-21水平明显下降(P<0.05)。结论 SYBR GreenⅠ实时荧光定量PCR是一种快速简便、敏感性高、特异性好的检测方法,对宫颈癌早期诊断、分子分期和预后判断有很好的应用前景。Objective To establish a method of SYBR greenⅠ real-time fluorescence quantitative PCR（FQ-PCR） for the determination of the expression levels of miR-21 in serum,and evaluate the significance of miR-21 in clinical monitoring of cervical diseases.Methods According to the sequence of miR-21 and U6,reverse-transcriptional primer for the stem-loop and the primers for real-time PCR were designed and synthesized.The expression levels of miR-21 in serum from 32 different cervical disease patients were determined and U6 was used as control.Results The experimental results of SYBR GreenⅠFQ-PCR with stem-loop RT primer were highly specific.The melting curve of miR-21 in serum showed a single peak,and the product of PCR was specific.High expression levels of miR-21 were detectable in the sera of cervical cancer patients,compared with those of uterine fibroid patients.The levels of miR-21 of cervical cancer patients obviously decreased after surgical resection（P〈0.05）.The difference showed a statistical significance.Conclusion The method of SYBR GreenⅠFQ-PCR with stem-loop RT primer is rapid,sensitive and specific to detect miR-21,which should be a promising early diagnostic approach for cervical cancer,and also its molecular staging and prognostic evaluation.

关 键 词：SYBRGreenⅠ实时荧光定量PCR MIR-21 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤]