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MicroRNA-221 regulates proliferation of bovine mammary gland epithelial cells by targeting the STAT5a and IRS1 genes

文献类型： 外文期刊

作者： Jiao, B. L. 1 ; Zhang, X. L. 1 ; Wang, S. H. 1 ; Wang, L. X. 1 ; Luo, Z. X. 1 ; Zhao, H. B. 2 ; Khatib, H. 3 ; Wang, X. 1 ;

作者机构： 1.Northwest A&F Univ, Coll Anim Sci & Technol, Yangling 712100, Shaanxi, Peoples R China

2.Shandong Acad Agr Sci, Inst Anim Sci & Vet Med, Jinan 250100, Shandong, Peoples R China

3.Univ Wisconsin, Dept Anim Sci, Madison, WI 53706 USA

关键词： miR-221; STAT5a; IRS1; bovine mammary gland epithelial cell; cell proliferation

期刊名称：JOURNAL OF DAIRY SCIENCE （ 影响因子：4.034； 五年影响因子：4.354 ）

ISSN： 0022-0302

年卷期： 2019 年 102 卷 1 期

页码：

收录情况： SCI

摘要： MicroRNAs (miRNA) play an essential role in mammary gland development and lactation. Previous studies in cattle have shown that miR-221 is highly expressed in peak compared with early lactation. However, the functions of miR-221 in bovine mammary gland epithelial cells and the mechanisms by which this miRNA affects cell proliferation and milk synthesis remain unclear. We hypothesized that miR-221 targets and modulates the expression of specific genes in the Janus kinase-signal transducer and activator of transcription (JAK-STAT) and phosphatidylinositol 3-kinase-proteinkinase B/ mammalian target of rapamycin (PI3K-Akt/mTOR) signaling pathways, which have crucial roles in lactation in cattle. Following transfection of rniR-221 into cultured bovine mammary gland epithelial cells, inhibition of cell proliferation and reduced viability of these cells were observed using the 3-(4,5-dimethylthiazol-2-yI)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis. To elucidate the molecular mechanisms of the effects of miR-221 on cell proliferation, we selected potential candidate genes that can be targeted by rniR-221 using bioinformatics prediction tools. The dual luciferase assay revealed that STAT5a, STATS, and IRS1 interact with miR-221 by its direct binding to the 3'-untranslated regions (UTR) of these genes. Subsequent analysis showed that transfection of a miR-221 mimic resulted in significantly decreased expression of STAT5a and IRS1 at both the RNA and protein levels using quantitative real-time PCR and Western blot analyses. Furthermore, expression levels of the downstream genes SOCS3, AKT3, and mTOR that are regulated by STAT5a and IRS1 in the JAK-STAT and PI3K-Akt/mTOR signaling pathways, were also altered after rniR-221 transfection. This is the first study to reveal the mechanisms by which miR-221 in-hibits mammary gland epithelial cell proliferation by targeting STAT5a and IRS1, key genes in the PI3K-Akt/mTOR and JAK-STAT signaling pathways.

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