miR-16-5p Promotes Erythroid Maturation of Erythroleukemia Cells by Regulating Ribosome Biogenesis

Abstract

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.

Insight into microRNAs’ involvement in hematopoiesis: current standing point of findings

“…Merkerova et al (2008) found that miR-16 was highly expressed in most human hematopoietic cells [ 6 ]. There are similar reports regarding miR-16-5p [ 66 ]. Although miR-16–2 is associated with polychromatic erythroblast differentiation, details of their role have not been established yet [ 4 ].…”

Comprehensive Analysis of microRNAs in Human Adult Erythropoiesis

“…* When the Ct values were undermined due to very low expression, a value of 40 was denoted for calculation. References [ 9 , 20 , 22 , 32 , 41 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 , 106 , 107 , 108 , 109 , 110 , 111 , 112 , 113 , 114 , 115 , 116 , 117 ] are referred to in the Supplementary Material file.…”

MicroRNA‐16‐5p exacerbates sepsis by upregulating aerobic glycolysis via SIRT3‐SDHA axis

“…By knockdown of miR‐16‐5p, we proved that miR‐16‐5p is essential for promoting aerobic glycolysis during sepsis. miR‐16‐5p directly binds to multiple target genes and modulates their expression (Cai et al, 2018; Papagiannopoulos et al, 2021; Qu et al, 2018; Ruan & Qian, 2019). However, its target in the macrophage, especially during infection, remains elusive.…”