荧光定量PCR检测宫颈癌患者血清中miR-203表达水平及意义

Serum miR-203 expression level detected by fluorescence quantitative PCR in cervical cancer patients and its significance

目的建立SYBR GreenⅠ实时荧光定量PCR检测血清中miR-203的表达水平,并检测宫颈癌患者、良性宫颈疾病及健康对照者血清miR-203的表达水平。方法设计miR-203、U6茎环逆转录引物和PCR扩增引物进行荧光定量PCR,以U6为内参相对定量比较不同宫颈疾病患者血清中miR-203的表达水平。结果该研究建立的方法能特异性检测到血清中miR-203的扩增信号,熔解曲线单一,PCR产物特异。宫颈癌患者血清中miR-203表达水平明显高于子宫肌瘤、宫颈炎等其他良性宫颈疾病,差异有统计学意义(P<0.05)。结论 SYBR GreenⅠ实时荧光定量PCR是一种快速简便、灵敏度高、特异度好的检测方法,可能在宫颈癌辅助诊断中有较好的应用前景。

Objective To establish the method of the SYBR GreenⅠreal-time fluorescent quantitative PCR for detecting the serum miR-203 expression level,and to detect the serum miR-203 expression levels in the patients with cervical cancer,cervical benign diseases and healthy controls.Methods The miR-203,U6 stem loop RT primers and the PCR amplification primers were designed for conducting fluorescence quantitative PCR,with U6 as the internal relative quantification,the serum miR-203 levels were compared among different cervical diseases.Results The established method could specifically detect the amplification signal of serum miR-203,the melting curve was single and PCR products were specific.The serum miR-203 level in the patients with cervical cancer was significantly higher than that in the patients with benign cervical diseases such as hysteromyoma and cervicitis,the difference was statistically significant（P〈0.05）.Conclusion The SYBR Green Ⅰ real-time fluorescent quantitative PCR is a quick,simple detection method with high sensitivity and good specificity,which may have a better application prospect in cervical cancer auxiliary diagnosis.