Liu Qingxin, Fan Jialong, Zhou Chuang, Wang Liqun, Zhao Bin, Zhang Haibin, Liu Bin, Tong Chunyi
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China.
Jiangsu Vocational College of Agriculture and Forestry, Jurong, Jiangsu 212400, China.
Int J Anal Chem. 2018 Sep 2;2018:3625823. doi: 10.1155/2018/3625823. eCollection 2018.
As a new tumor marker, the microRNA-21 (miRNA21) level can provide important information for early diagnosis, drug treatment, and prognosis of gastric cancer. With the tool of molecular beacons which can hybridize specifically with target miRNA-21 and generate fluorescence signal change, this paper develops a direct, simple, and rapid method for miRNA-21 detection with detection limit of 0.5 nM. Under the optimal conditions, the method was used to detect the expression of miRNA-21 in tumor cells and tissues. The results showed significant differences of miRNA-21 levels in tumor cells which have different origins and different degree of malignancy. In 8 cases of gastric cancer tissues and adjacent tissues, the level of miRNA-21 in 6 cases was higher than that in adjacent tissues, 1 case had lower expression level than that in adjacent tissues, and 1 case had no significant difference. Furthermore, qRT-PCR method was used to verify the detection results based on the fluorescent probe detection method. The consistent results show that the molecular beacon assay has a good prospect in direct and rapid detection of miRNA-21 expression and will be widely used in the functional research and clinical diagnosis of microRNA.
作为一种新型肿瘤标志物,微小RNA-21(miRNA21)水平可为胃癌的早期诊断、药物治疗及预后提供重要信息。利用能与靶标miRNA-21特异性杂交并产生荧光信号变化的分子信标工具,本文开发了一种直接、简单且快速的miRNA-21检测方法,检测限为0.5 nM。在最佳条件下,该方法用于检测肿瘤细胞和组织中miRNA-21的表达。结果显示,不同来源和不同恶性程度的肿瘤细胞中miRNA-21水平存在显著差异。在8例胃癌组织及癌旁组织中,6例miRNA-21水平高于癌旁组织,1例表达水平低于癌旁组织,1例无显著差异。此外,采用qRT-PCR方法对基于荧光探针检测法的检测结果进行验证。一致的结果表明,分子信标分析法在直接快速检测miRNA-21表达方面具有良好前景,将广泛应用于微小RNA的功能研究和临床诊断。