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目的神经胶质瘤是中枢神经系统中最普遍、最具侵袭性的原发性肿瘤,具有高发病率和高死亡率的特征。microRNAs(miRNAs)是一类平均长约22nt的非编码小RNA,主要在转录后水平负责调控基因的表达,参与癌症发生发展过程。miR-23a起着原癌基因的作用,在多种恶性肿瘤中miR-23a高表达,本文比较人正常脑组织和胶质瘤细胞系中miR-23a的相对表达水平。方法分别抽提正常脑组织和胶质瘤细胞系中的总RNA,设计反转录miR-23a的茎环引物及定量的正反向引物,使用SYBR Green试剂对其进行实时荧光定量PCR(qRT-PCR)检测。结果设计的引物能高效且特异地逆转录并且扩增miR-23a;miR-23a在胶质瘤细胞系A172、U87、U251中均显著高表达,是正常脑组织表达量的4倍以上。结论设计的miR-23a引物能有效地进行qRT-PCR;miR-23a在胶质瘤细胞系中高表达。
Objective Glioma is the most common and aggressive primary tumor in the central nervous system with high morbidity and mortality. MicroRNAs (miRNAs) are a class of non-coding small RNAs with an average length of 22 nt. They are mainly responsible for the regulation of gene expression at the post-transcriptional level and are involved in the development of cancer. miR-23a plays a role of proto-oncogene and miR-23a is highly expressed in a variety of malignant tumors. In this paper, the relative expression levels of miR-23a in normal human brain and glioma cell lines were compared. Methods The total RNA was extracted from normal brain tissue and glioma cell lines respectively. The stem-loop primers and quantitative forward-reverse primers of miR-23a were designed and reverse transcribed. SYBR Green reagent was used to perform real-time fluorescence quantitative PCR (qRT -PCR) assay. Results The designed primers could reverse and efficiently reverse miR-23a expression. MiR-23a was highly expressed in glioma cell lines A172, U87 and U251, which was more than four times higher than normal brain tissue. Conclusion The designed miR-23a primer can effectively carry out qRT-PCR; miR-23a is highly expressed in glioma cell lines.