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  

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

作　　者：郭文涛[1] 畅灵丽[1] 赵青[1] 张清伟[2] 朱利伟[3] 李亚光[1] 孙宁[4] 张威[5] 

机构地区：[1]漯河医学高等专科学校基础医学部病原生物学与免疫学教研室,河南漯河462002 [2]漯河医学高等专科学校第一附属医院妇产科,河南漯河462002 [3]漯河医学高等专科学校第三附属医院外科,河南漯河462002 [4]南京军区南京总医院临床中心实验科,南京210002 [5]郑州大学第三附属医学病理科,郑州450000

出　　处：《广东医学》2016年第6期837-840,共4页Guangdong Medical Journal

基　　金：漯河医学高等专科学校研究计划项目(编号:2013-S-LMC21)

摘　　要：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。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 extraction was detected by Bland- Altman method. Utility of miR- 21 expression detected by MS in screening ESCC was evaluated 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 expression 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 expression 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.

关 键 词：磁性纳米微粒 实时荧光RT-PCR MIR-21 食管鳞状细胞癌 

分 类 号：R373.13[医药卫生—病原生物学]