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14-3-3γ affects mTOR pathway and regulates lactogenesis in dairy cow mammary epithelial cells

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Abstract

14-3-3 proteins are an acidic protein family that is highly conserved and widely distributed in eukaryotic cells. Recent studies have found that 14-3-3 proteins play critical roles in cell signal transductions, cell growth and differentiation, and protein synthesis. 14-3-3γ is an important member of 14-3-3 protein family. In our previous study, we found that 14-3-3γ was upregulated by estrogen in dairy cow mammary epithelial cell (DCMEC), but the function and mechanism of 14-3-3γ is not known. In this experiment, we first cultured and purified the primary DCMEC and found 14-3-3γ located both in the cytoplasm and nucleus by using immunofluorescence assay. Methionine, lysine, estrogen, and prolactin could upregulate the expression of 14-3-3γ, stimulate the secretion of β-casein and triglyceride, and raise the cell viability of DCMEC. We constructed a stable 14-3-3γ overexpression cell line of DCMEC and found that the expressions of mTOR and p-mTOR, the secretion of triglyceride and β-casein (CSN2), and the cell viability of DCMEC were all upregulated. We also observed the effects of 14-3-3γ gene silencing and gained consistent results with 14-3-3γ overexpression. These findings reveal that 14-3-3γ affects the mTOR pathway and regulates lactogenesis in DCMECs.

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Acknowledgments

This study was financially supported by 863 Project of Ministry of Science and Technology of China (No: 2013AA102504-03) and the Major State Basic Research Development Program of China (973 Program, 2011CB100804).

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Authors and Affiliations

  1. Key Laboratory of Agricultural Biological Functional Genes, Northeast Agricultural University, Harbin, 150030, China

    Nagam Khudhair, Chaochao Luo, Li Zhang, Shuang Zhang, Jinxia Ao, Qingzhang Li & Xuejun Gao

  2. College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, China

    Ahmed Khalid

  3. Biology Department, Education College for Women, Al-Anbar University, Ramadi, 31001, Iraq

    Nagam Khudhair

Authors
  1. Nagam Khudhair
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  2. Chaochao Luo
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  3. Ahmed Khalid
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  4. Li Zhang
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  5. Shuang Zhang
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  6. Jinxia Ao
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  7. Qingzhang Li
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  8. Xuejun Gao
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Corresponding author

Correspondence to Xuejun Gao.

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Editor: T. Okamoto

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Khudhair, N., Luo, C., Khalid, A. et al. 14-3-3γ affects mTOR pathway and regulates lactogenesis in dairy cow mammary epithelial cells. In Vitro Cell.Dev.Biol.-Animal 51, 697–704 (2015). https://doi.org/10.1007/s11626-015-9879-x

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  • DOI: https://doi.org/10.1007/s11626-015-9879-x

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