Nedaeinia Reza, Sharifi Mohammadreza, Avan Amir, Kazemi Mohammad, Nabinejad Abdolreza, Ferns Gordon A, Ghayour-Mobarhan Majid, Salehi Rasoul
1 Department of Medical Biotechnology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
2 Students Research Committee, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Tumour Biol. 2017 Mar;39(3):1010428317692261. doi: 10.1177/1010428317692261.
Colorectal cancer is among the most lethal of malignancies, due to its propensity to metastatic spread and multifactorial-chemoresistance. The latter property supports the need to identify novel therapeutic approaches for the treatment of colorectal cancer. MicroRNAs are endogenous non-coding small RNA molecules that function as post-transcriptional regulators of gene expression. Recently, programmed cell death 4 has been identified as a protein that increases during apoptosis. This gene is among the potential targets of miR-21 (OncomiR). Locked nucleic acid-modified oligonucleotides have recently emerged as a potential therapeutic option for targeting microRNAs. The aim of this study was to explore the functional role of locked nucleic acid-anti-miR-21 in the LS174T cell line in vitro and in vivo models. LS174T cells were treated with locked nucleic acid-anti-miR-21 for 24, 48, and 72 h in vitro. The expression of miR-21 and PDCD4 at messenger RNA (mRNA) level was evaluated by quantitative real-time polymerase chain reaction, while the protein level of PDCD4 was determined by Western blotting. Cell migratory behavior and the cluster-forming ability of cells were assessed before and after therapy. The disseminated tumor cells were assessed in the chick chorioallantoic membrane model by Alu quantitative polymerase chain reaction. Locked nucleic acid-anti-miR-21 was transfected successfully into the LS174T cells and inhibited the expression of miR-21. Locked nucleic acid-anti-miR-21 inhibited the migration and the number of cells forming clusters. Moreover, we found that locked nucleic acid-anti-miR-21 transfection was associated with a significant reduction in metastatic properties as assessed by the in ovo model. Our findings demonstrated the novel therapeutic potential of locked nucleic acid-anti-miR-21 in colon adenocarcinoma with high miR-21 expression.
由于结直肠癌易于发生转移扩散且具有多因素化学抗性，它是最致命的恶性肿瘤之一。后一种特性支持了识别用于治疗结直肠癌的新治疗方法的必要性。微小RNA是内源性非编码小RNA分子，其作为基因表达的转录后调节因子发挥作用。最近，程序性细胞死亡4已被鉴定为一种在细胞凋亡期间增加的蛋白质。该基因是miR-21（致癌miRNA）的潜在靶标之一。锁定核酸修饰的寡核苷酸最近已成为靶向微小RNA的潜在治疗选择。本研究的目的是在体外和体内模型中探索锁定核酸抗miR-21在LS174T细胞系中的功能作用。在体外，用锁定核酸抗miR-21处理LS174T细胞24、48和72小时。通过定量实时聚合酶链反应评估miR-21和程序性细胞死亡4在信使RNA（mRNA）水平的表达，而通过蛋白质印迹法测定程序性细胞死亡4的蛋白质水平。在治疗前后评估细胞的迁移行为和细胞的集落形成能力。通过Alu定量聚合酶链反应在鸡胚绒毛尿囊膜模型中评估播散的肿瘤细胞。锁定核酸抗miR-21成功转染到LS174T细胞中并抑制了miR-21的表达。锁定核酸抗miR-21抑制了迁移和形成集落的细胞数量。此外，我们发现，通过卵内模型评估，锁定核酸抗miR-2转染与转移特性的显著降低相关。我们的研究结果证明了锁定核酸抗miR-21在具有高miR-21表达的结肠腺癌中的新治疗潜力。
