Abstract
The development of the udder and the milk yield are closely related to the number and vitality of mammary epithelial cells. Many previous studies have proved that non-coding RNAs (ncRNAs) are widely involved in mammary gland development and the physiological activities of lactation. Our laboratory previous sequencing data revealed that miR-574-5p was differentially expressed during the colostrum and peak lactation stages, while the molecular mechanism of the regulatory effect of miR-574-5p on goat mammary epithelial cells (GMECs) is unclear. In this study, the targeting relationship was detected between miR-574-5p or ecotropic viral integration site 5-like (EVI5L) and circRNA-006258. The results declared that miR-574-5p induced the down-regulation of EVI5L expression at both the mRNA and protein levels, while circRNA-006258 relieved the inhibitory effect through adsorbing miR-574-5p. EVI5L blocked the G1 phase and promoted the S phase by activating the Rab23/ITGB1/TIAM1/Rac1-TGF-β/Smad pathway in GMECs. By increasing the protein expression of Bcl2 and reducing the protein expression of Bax, EVI5L promoted cell growth and inhibited apoptosis. The activation of the PI3K/AKT–mTOR signaling pathway promoted the production of triacylglycerol (TAG) and β-casein in GMECs. The circRNA–006258/miR-574-5p/EVI5L axis could regulate the cell growth and milk synthesis of GMECs by sponge-adsorbed miR-574-5p. These results would provide scientific evidence for precision animal breeding in the industry of dairy goats.
Highlights
MicroRNAs are a series of small non-coding RNA gene products containing 18–22 nucleotides
The 3’ untranslated region (3’ UTR) of ecotropic viral integration site 5-like (EVI5L) has the special nucleotide sequence which could be attached to miR-574-5p (Figure 1A,B)
There was no significant change in the co-transcriptional group of miR-574-5p and MUT-EVI5L-psiCHECK2
Summary
MicroRNAs (miRNAs) are a series of small non-coding RNA gene products containing 18–22 nucleotides. Of target messenger RNAs (mRNAs) and negatively regulate gene expression by transcriptional or translational inhibition [1,2,3]. MiR-574-5p are implicated in the pathophysiology of many tumors, including breast cancer, colorectal cancer and other oncoma [4,5,6]. MiR-574-5p is a very important regulator for cell migration and proliferation [7,8,9]. Based on our laboratory sequencing data, miR-574-5p is differentially expressed in the colostrum and peak lactation periods [10]. MiR-574-5p targets MAP3K9 and reduces the secretion of β-casein and triglycerides through PI3K/AKT–mTOR pathways in goat mammary epithelial cells (GMECs) [11]
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