Papagiannopoulos Christos I, Theodoroula Nikoleta F, Vizirianakis Ioannis S
Laboratory of Pharmacology, School of Pharmacy, Aristotle University of Thessaloniki, GR-54124 Thessaloniki, Greece.
FunPATH (Functional Proteomics and Systems Biology Research Group at AUTH) Research Group, KEDEK-Aristotle University of Thessaloniki, Balkan Center, GR-57001 Thessaloniki, Greece.
Pharmaceuticals (Basel). 2021 Feb 9;14(2):137. doi: 10.3390/ph14020137.
miRNAs constitute a class of non-coding RNA that act as powerful epigenetic regulators in animal and plant cells. In order to identify putative tumor-suppressor miRNAs we profiled the expression of various miRNAs during differentiation of erythroleukemia cells. RNA was purified before and after differentiation induction and subjected to quantitative RT-PCR. The majority of the miRNAs tested were found upregulated in differentiated cells with miR-16-5p showing the most significant increase. Functional studies using gain- and loss-of-function constructs proposed that miR-16-5p has a role in promoting the erythroid differentiation program of murine erythroleukemia (MEL) cells. In order to identify the underlying mechanism of action, we utilized bioinformatic in-silico platforms that incorporate predictions for the genes targeted by miR-16-5p. Interestingly, ribosome constituents, as well as ribosome biogenesis factors, were overrepresented among the miR-16-5p predicted gene targets. Accordingly, biochemical experiments showed that, indeed, miR-16-5p could modulate the levels of independent ribosomal proteins, and the overall ribosomal levels in cultured cells. In conclusion, miR-16-5p is identified as a differentiation-promoting agent in erythroleukemia cells, demonstrating antiproliferative activity, likely as a result of its ability to target the ribosomal machinery and restore any imbalanced activity imposed by the malignancy and the blockade of differentiation.
微小RNA（miRNA）构成了一类非编码RNA，在动植物细胞中作为强大的表观遗传调节因子发挥作用。为了鉴定潜在的肿瘤抑制性miRNA，我们分析了红白血病细胞分化过程中各种miRNA的表达情况。在诱导分化前后纯化RNA，并进行定量逆转录聚合酶链反应（RT-PCR）。测试的大多数miRNA在分化细胞中上调，其中miR-16-5p的增加最为显著。使用功能获得和功能丧失构建体进行的功能研究表明，miR-16-5p在促进小鼠红白血病（MEL）细胞的红系分化程序中发挥作用。为了确定其潜在的作用机制，我们利用了生物信息学的计算机平台，该平台整合了对miR-16-5p靶向基因的预测。有趣的是，核糖体成分以及核糖体生物发生因子在miR-16-5p预测的基因靶标中占比过高。相应地，生化实验表明，事实上，miR-16-5p可以调节培养细胞中独立核糖体蛋白的水平以及整体核糖体水平。总之，miR-16-5p被鉴定为红白血病细胞中的一种促分化因子，具有抗增殖活性，这可能是由于它能够靶向核糖体机制并恢复由恶性肿瘤和分化阻滞所导致的任何失衡活性。
