miR-380-3p promotes β-casein expression by targeting αS1-casein in goat mammary epithelial cells

Song, Ning; Luo, Jun; Huang, Lian; Chen, Xiaoying; Niu, Huimin; Zhu, Lu

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Anim Biosci > Accepted Articles

Article

https://doi.org/10.5713/ab.23.0007 [Accepted] Published online May 4, 2023.

miR-380-3p promotes β-casein expression by targeting αS1-casein in goat mammary epithelial cells

Ning Song^1,2

, Jun Luo^1,*

¹Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China
²College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China

Correspondence: Jun Luo, Tel: +86-029-87082891, Fax: +86-029-87082892, Email: luojun@nwafu.edu.cn

Received: 5 January 2023 • Revised: 23 February 2023 • Accepted: 28 March 2023

Abstract

Objective
αS1-Casein is more closely associated with milk allergic reaction than other milk protein components. microRNA (miRNA) is a class of small non-coding RNAs that modulate multiple biological progresses by the target gene. However, the post-transcriptional regulation of αS1-casein expression by miRNA in ruminants remains unclear. This study aims to explore the regulatory roles of miR-380-3p on αS1-casein synthesis in goat mammary epithelial cells (GMEC).
Methods
αS1-Casein gene and miR-380-3p expression was measured in dairy goat mammary gland by quantitative real-time PCR (qRT-PCR). miR-380-3p overexpression and knockdown were performed by miR-380-3p mimic or inhibitor in GMEC. The effect of miR-380-3p on αS1-casein synthesis was detected by qRT-PCR, western blot, luciferase and chromatin immunoprecipitation assays in GMEC.
Results
Compared with middle-lactation period, αS1-casein gene expression is increased, while miR-380-3p expression is decreased during peak-lactation of dairy goats. miR-380-3p reduces αS1-casein abundance by targeting the 3’-untranslated region (3’UTR) of αS1-casein mRNA in GMEC. miR-380-3p enhances β-casein expression and signal transducer and activator of transcription 5a (STAT5a) activity. Moreover, miR-380-3p promotes β-casein abundance through target gene αS1-casein, and activates β-casein transcription by enhancing the binding of STAT5 to β-casein gene promoter region.
Conclusion
miR-380-3p decreases αS1-casein expression and increases β-casein expression by targeting αS1-casein in GMEC, which supplies a novel strategy for reducing milk allergic potential and building up milk quality in ruminants.

Keywords: αS1-Casein; Goat Mammary Epithelial Cells; Milk Allergy; miR-380-3p