Effects of Different Concentrations of Linoleic Acid on Expression of Milk Protein Synthesis Related Gene in Two-Dimension and Three-Dimension Cultured Bovine Mammary Epithelial Cells

XING Yuanyuan; LI Dabiao; LI Zinan; JIN Yaya

doi:10.3969/j.issn.1006-267x.2019.10.031

2019 , Vol. 31 >Issue 10: 4667 - 4674

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.10.031

Effects of Different Concentrations of Linoleic Acid on Expression of Milk Protein Synthesis Related Gene in Two-Dimension and Three-Dimension Cultured Bovine Mammary Epithelial Cells

XING Yuanyuan ,
LI Dabiao ,
LI Zinan ,
JIN Yaya

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College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2019-04-06

Online published: 2019-10-17

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Abstract

This study aims to evaluate the effects different concentrations of linoleic acid (LA) on expression of milk protein synthesis related gene in two-dimension (2D) and three-dimension (3D) cultured bovine mammary epithelial cells (BMECs). A single factorial experimental design was used in the experiment, the P3 generation BMECs were seeded in 6-well plates with Matrigel spread, and cultured for 3 d, next, different concentrations (0, 40 and 80 μmol/L) of LA were supplemented in the culture medium for 48 h. The same concentration of LA was supplemented in BMECs in 2D and 3D cultured models, to compare the cell morphology in BMECs in 2D and 3D cultured models, and the expression of milk protein synthesis related genes were determined by real-time fluorescence quantitative method. The results showed as follow:1) BMECs showed cobblestone morphology in 2D culture model and alveoli-like structures in 3D culture model. 2) The expression of milk protein synthesis related genes in BMECs in 2D culture model was significantly higher than that in 3D culture model (P<0.05). 3) The expression of α-casein (CSN1S1), signal transduction activator 5 (STAT5A), eukaryotic translation initiation factor 4E binding protein 1 (EIF4EBP1) and ribosomal p70s6 kinase (RPS6K1) in 2D and 3D culture models were the same, however, the expression of κ-casein (CSN3), mammalian target of rapamycin (mTOR) and serine/threonine kinase (AKT1) in 2D and 3D culture models were showed an opposite trend. To sum up, 3D culture model is a more appropriate model for the study related to CSN3, mTOR and AKT1 genes expression.

Key words： dairy cows; three-dimension cell culture model; linoleic acid; milk protein synthesis

Cite this article

XING Yuanyuan , LI Dabiao , LI Zinan , JIN Yaya . Effects of Different Concentrations of Linoleic Acid on Expression of Milk Protein Synthesis Related Gene in Two-Dimension and Three-Dimension Cultured Bovine Mammary Epithelial Cells[J]. Chinese Journal of Animal Nutrition, 2019 , 31(10) : 4667 -4674 . DOI: 10.3969/j.issn.1006-267x.2019.10.031

References

[1] LOCK A L,BAUMAN D E.Modifying milk fat composition of dairy cows to enhance fatty acids beneficial to human health[J].Lipids,2004,39(12):1197-1206.

[2] REYES N,REYES I,TIWARI R,et al.Effect of linoleic acid on proliferation and gene expression in the breast cancer cell line T47D[J].Cancer Letters,2004,209(1):25-35.

[3] MACH N,JACOBS A A A,KRUIJT L,et al.Alteration of gene expression in mammary gland tissue of dairy cows in response to dietary unsaturated fatty acids[J].Animal,2011,5(8):1217-1230.

[4] CANT J P,DEPETERS E J,BALDWIN R L.Mammary amino acid utilization in dairy cows fed fat and its relationship to milk protein depression[J].Journal of Dairy Science,1993,76(3):762-774.

[5] HE M,ARMENTANO L E.Effect of fatty acid profile in vegetable oils and antioxidant supplementation on dairy cattle performance and milk fat depression[J].Journal of Dairy Science,2011,94(5):2481-2491.

[6] KADEGOWDA A K G,PIPEROVA L S,DELMONTE P,et al.Abomasal infusion of butterfat increases milk fat in lactating dairy cows[J].Journal of Dairy Science,2008,91(6):2370-2379.

[7] YONEZAWA T,YONEKURA S,KOBAYASHI Y,et al.Effects of long-chain fatty acids on cytosolic triacylglycerol accumulation and lipid droplet formation in primary cultured bovine mammary epithelial cells[J].Journal of Dairy Science,2004,87(8):2527-2534.

[8] YAMADA K M,CUKIERMAN E.Modeling tissue morphogenesis and cancer in 3D[J].Cell,2007,130(4):601-610.

[9] HSIAO A Y,TUNG Y C,KUO C H,et al.Micro-ring structures stabilize microdroplets to enable long term spheroid culture in 384 hanging drop array plates[J].Biomedical Microdevices,2012,14(3):313-323.
[10] DEBNATH J,BRUGGE J S.Modelling glandular epithelial cancers in three-dimensional cultures[J].Nature Reviews Cancer,2005,5(9):675-688.

[11] MURTAGH J,MCARDLE E,GILLIGAN E,et al.Organization of mammary epithelial cells into 3D acinar structures requires glucocorticoid and JNK signaling[J].The Journal of Cell Biology,2004,166(1):133-143.

[12] YONEZAWA T,HAGA S,KOBAYASHI Y,et al.Unsaturated fatty acids promote proliferation via ERK1/2 and Akt pathway in bovine mammary epithelial cells[J].Biochemical and Biophysical Research Communications,2008,367(4):729-735.

[13] 王秀美.二、三维培养模式对奶牛乳腺上皮细胞泌乳代谢相关基因的影响[D].博士学位论文.呼和浩特:内蒙古农业大学,2013.
[14] KOZLOWSKI M,WILCZAK J,MOTYL T,et al.Role of extracellular matrix and prolactin in functional differentiation of bovine BME-UV1 mammary epithelial cells[J].Polish Journal of Veterinary Sciences,2001,14(3):3433-3442.
[15] SCHMEICHEL K L,BISSELL M J.Modelling tissue-specific signaling and organ function in three dimensions[J].Journal of Cell Science,2003,116(12):2377-2388.

[16] SASSER A K,MUNDY B L,SMITH K M,et al.Human bone marrow stromal cells enhance breast cancer cell growth rates in a cell line-dependent manner when evaluated in 3D tumor environments[J].Cancer Letters,2007,254(2):255-264.

[17] BISSELL M J,AGGELER J.Dynamic reciprocity:how do extracellular matrix and hormones direct gene expression?[J].Progress in Clinical and Biological Research 1987,249:251-262.
[18] BISSELL M J,HALL H G,PARRY G.How does the extracellular matrix direct gene expression?[J].Journal of Theoretical Biology,1982,99(1):31-68.

[19] LEE G Y,KENNY P A,LEE E H,et al.Three-dimensional culture models of normal and malignant breast epithelial cells[J].Nature Methods,2007,4(4):359-365.

[20] YANG X,YANG C,FARBERMAN A,et al.The mammalian target of rapamycin-signaling pathway in regulating metabolism and growth[J].Journal of Animal Science,2008,86(Suppl.14):E35-E50.
[21] SHENG R,YAN S M,QI L Z,et al.Effect of the ratios of unsaturated fatty acids on the expressions of genes related to fat and protein in the bovine mammary epithelial cells[J].In vitro Cellular & Developmental Biology-Animal,2015,51(4):381-389.

[22] PORSTMANN T,SANTOS C R,GRIFFITHS B,et al.SREBP activity is regulated by mTORCl and contributes to Akt-dependent cell growth[J].Cell Metabolism,2008,8(3):224-236.

[23] BURGOS S A,DAI M,CANT J P.Nutrient availability and lactogenic hormones regulate mammary protein synthesis through the mammalian target of rapamycin signaling pathway[J].Journal of Dairy Science,2010,93(1):153-161.

[24] KIM J E,CHEN J.Regulation of peroxisome proliferator-activated receptor-γ activity by mammalian target of rapamycin and amino acids in adipogenesis[J].Diabetes,2004,53(11):2748-2756.

[25] HADLEY M A,BYERS S W,SUÁREZ-QUIAN C A,et al.Extracellular matrix regulates Sertoli cell differentiation,testicular cord formation,and germ cell development in vitro[J].Journal of Cell Biology,1985,101(4):1511-1522.

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