Abstract
In a previous study, we found miR-10b to be more abundant in a conditioned culture medium of degenerate embryos compared to that of blastocysts. Here, we show that miR-10b mimics added to the culture medium can be taken up by embryos. This uptake results in an increase in embryonic cell apoptosis and aberrant expression of DNA methyltransferases (DNMTs). Using several algorithms, Homeobox A1 (HOXA1) was identified as one of the potential miR-10b target genes and dual-luciferase assay confirmed HOXA1 as a direct target of miR-10b. Microinjection of si-HOXA1 into embryos also resulted in an increase in embryonic cell apoptosis and downregulation of DNMTs. Cell progression analysis using Madin–Darby bovine kidney cells (MDBKs) showed that miR-10b overexpression and HOXA1 knockdown results in suppressed cell cycle progression and decreased cell viability. Overall, this work demonstrates that miR-10b negatively influences embryo quality and might do this through targeting HOXA1 and/or influencing DNA methylation.
Highlights
MiRNAs, small non-coding RNAs, function as crucial regulators that can be transferred between cells (Valadi et al, 2007)
Several studies have shown that miR-10b plays important roles in cell apoptosis, cell proliferation, cell migration, and invasion in miR-10b Negatively Influences Embryos Quality human cancer cells (Wang et al, 2007; Liao et al, 2014; Chen et al, 2016; Zhen et al, 2016; Zhu et al, 2016; Guan et al, 2018), mouse cells (Tan et al, 2018), and goat granulosa cells (Peng et al, 2016)
As one of the HOX family members, Homeobox A1 (HOXA1) is involved in various biological processes, including cell apoptosis and growth (Zhang et al, 2018)
Summary
MiRNAs, small non-coding RNAs, function as crucial (epigenetic) regulators that can be transferred between cells (Valadi et al, 2007). We identified 114 known and 180 novel secreted miRNAs present in bovine embryo culture media (CM). Of these miRNAs, miR-30c and miR-10b were much more abundant in CM of slow-cleaving embryos compared to intermediate-cleaving embryos. MiR-10b was shown to be more abundant in the culture medium of degenerate embryos compared with that of blastocysts, and more abundant in the culture medium of slow-cleaving embryos compared with that of intermediate-cleaving embryos, indicating that overexpression of miR-10b has a negative influence on preimplantation embryo development in cattle (Lin et al, 2019). MiR-10b has been shown to regulate cell miR-10b Negatively Influences Embryos Quality invasion, apoptosis, viability, and migration in multiple cell lines in human, mouse, and goat (Chen et al, 2016; Li et al, 2016; Peng et al, 2016; Zhen et al, 2016; Zhu et al, 2016; Tan et al, 2018)
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