SNAT2介导蛋氨酸对牛乳腺上皮细胞增殖与自噬的调节作用

doi:10.16431/j.cnki.1671-7236.2019.09.003

Abstract

Abstract: The sodium-dependent neutral amino acid transporter 2 (SNAT2) is an amino acid transporter which can transport neutral amino acids and is widely distributed in various cells.Amino acids is not only substrates for protein synthesis,but also key signal molecules regulating cell metabolism.However,it is little known whether SNAT2 mediates the regulation of amino acids on autophagy.In this study,CASY cell count and Western blotting were used to determine the proliferation of BMECs after SNAT2 overexpression and siRNA interference.The effects of SNAT2 on the expression of autophagic biomarker LC3-Ⅱ was detected by Western blotting and the change of autophagic spots (LC3-Ⅱ) in cells was determined by immunofluorescence.The results showed that SNAT2 overexpression increased the expression levels of p-PI3K,p-mTOR and cyclin D1,whereas SNAT2 knockdown had the opposite effects.Furthermore,the expression of LC3-Ⅱ and the autophagy spots were both increased in SNAT2-downregulated cells.The p-mTOR expression was increased and LC3-Ⅱ was decreased in cells treated with the autophagy enhancer trehalose dihydrate (Tre) together with methionine (Met) compared with the control (cells treated with Tre).The expression of p-mTOR was decreased and LC3-Ⅱ was increased in cells treated with Tre together with Met addition and transfected with a SNAT2 siRNA compared with the control (cells treated with Tre and Met).In conclusion,these data demonstrated that SNAT2 mediated the regulation of Met on proliferation of and autophagy in BMECs via PI3K-mTOR/Cyclin D1 signaling pathway.

Key words: SNAT2; BMECs; mTOR; proliferation; autophagy

CLC Number:

Q291

QI Hao, MENG Chunyu, GAO Xuejun, ZHEN Zhen. SNAT2: A Key Mediator for Methionine to Regulate Proliferation and Autophagy in Bovine Mammary Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(9): 2516-2525.

References

[1] YAO D,MACKENZIE B,MING H,et al.A novel system A isoform mediating Na⁺/neutral amino acid cotransport[J].Journal of Biological Chemistry,2000,275(30):22790-22797.
[2] CAPUCO A V,WOOD D L,BALDWIN R,et al.Mammary cell number,proliferation,and apoptosis during a bovine lactation:Relation to milk production and effect of bST[J].Journal of Dairy Science,2001,84(10):2177-2187.
[3] CHEN J,BAI M,NING C,et al.Gankyrin facilitates follicle-stimulating hormone-driven ovarian cancer cell proliferation through the PI3K/AKT/HIF-1α/cyclin D1 pathway[J].Oncogene,2015,35(19):2506-2517.
[4] MIZUNOE Y,WATANABE S,SUDO Y,et al.Trehalose protects against oxidative stress by regulating the Keap1-Nrf2 and autophagy pathways[J].Redox Biology,2018,15:115-124.
[5] MORTIMORE G E,POSO A R.Intracellular protein catabolism and its control during nutrient deprivation and supply[J].Annual Review of Nutrition,1987,7(1):539-564.
[6] SHI H,ZHAO X,DING Z,et al.Na⁺/H⁺ exchanger regulates amino acid-mediated autophagy in intestinal epithelial cells[J].Cellular Physiology and Biochemistry,2017,42(6):2418-2429.
[7] TAN H W S,SIM A Y L,LONG Y C.Glutamine metabolism regulates autophagy-dependent mTORC1 reactivation during amino acid starvation[J].Nature Communications,2017,8(1):338.
[8] SAXTON R A,SABATINI D M.mTOR Signaling in growth,metabolism,and disease[J].Cell,2017,169(2):361-371.
[9] GAO H N,ZHAO S G,ZHENG N,et al.Combination of histidine,lysine,methionine,and leucine promotes beta-casein synthesis via the mechanistic target of rapamycin signaling pathway in bovine mammary epithelial cells[J].Journal of Dairy Science,2017,100(9):7696-7709.
[10] ZHANG M,CHEN D,ZHEN Z,et al.Annexin A2 positively regulates milk synthesis and proliferation of bovine mammary epithelial cells through the mTOR signaling pathway[J].Journal of Cellular Physiology,2018,233(3):2464-2475.
[11] YU M,LUO C,HUANG X,et al.Amino acids stimulate glycyl-tRNA synthetase nuclear localization for mammalian target of rapamycin expression in bovine mammary epithelial cells[J].Journal of Cellular Physiology,2019,234(5):7608-7621.
[12] 孟春雨,黄鑫,刘丽杰,等.牛乳腺上皮细胞SNAT2对氨基酸调节乳合成的影响[J].中国畜牧兽医,2018,45(8):2119-2127. MENG C Y,HUANG X,LIU L J,et al.Effects of SNAT2 on amino acid-stimulated milk protein and milk fat synthesis in bovine mammary epithelial cells[J].China Animal Husbandry & Veterinary Medicine, 2018,45(8):2119-2127.(in Chinese)
[13] 史琳琳,赵锋,高学军,等.催乳复合物、胰岛素样生长因子-Ⅰ对奶牛乳腺上皮细胞中AKT-1和mTOR表达的作用[J].中国畜牧兽医,2013,40(3):23-27. SHI L L,ZHAO F,GAO X J,et al.Effection of DIP and IGF-Ⅰ on the expression of AKT-1 and mTOR genes in bovine mammary epithelial cells[J].China Animal Husbandry & Veterinary Medicine,2013,40(3):23-27.(in Chinese)
[14] 陈建晖,佟慧丽,李庆章,等.奶牛乳腺上皮细胞系的建立[J].畜牧兽医学报,2009,40(5):743-747. CHEN J H,TONG H L,LI Q Z,et al.Establishment of dairy cow mammary gland epithelial cell line[J].Chinese Journal of Animal and Veterinary Sciences,2009,40(5):743-747.(in Chinese)
[15] 李冬,黄鑫,李姗姗,等.METTL3对乳腺上皮细胞中乳蛋白和乳脂肪合成的影响[J].中国畜牧兽医,2017,44(5):1267-1274. LI D,HUANG X,LI S S,et al.Effect of METTL3 on milk protein and milk fat synthesis of bovine mammary epithelial cells[J].China Animal Husbandry & Veterinary Medicine,2017,44(5):1267-1274.(in Chinese)
[16] PINILLA J,ALEDO J C,CWIKLINSKI E,et al.SNAT2 transceptor signalling via mTOR:A role in cell growth and proliferation?[J].Frontiers in Bioscience,2011,3(4):1289-1299.
[17] KATE E,ZEERAK N,JEREMY B,et al.Inhibition of SNAT2 by metabolic acidosis enhances proteolysis in skeletal muscle[J].Journal of the American Society of Nephrology,2008,19(11):2119-2129.
[18] TANG Y,TAN B,LI G,et al.The regulatory role of MeAIB in protein metabolism and the mTOR signaling pathway in porcine enterocytes[J].International Journal of Molecular Sciences,2018,19(3):714.
[19] HASSANIAN S M,ARDESHIRYLAJIMI A,DINARVAND P,et al.Inorganic polyphosphate promotes cyclin D1 synthesis through activation of mTOR/Wnt/β-catenin signaling in endothelial cells[J].Journal of Thrombosis & Haemostasis,2016,14(11):2261-2273.
[20] WANG Q,SHI W.UNBS5162 inhibits SKOV3 ovarian cancer cell proliferation by regulating the PI3K/AKT signalling pathway[J].Oncology Letters,2019,17(3):2976-2982.
[21] QI H,MENG C,JIN X,et al.Methionine promotes milk protein and fat synthesis and cell proliferation via the SNAT2-PI3K signaling pathway in bovine mammary epithelial cells[J].Journal of Agricultural and Food Chemistry,2018,66(42):11027-11033.
[22] MAYER A L,HIGGINS C B,HEITMEIER M R,et al.SLC2A8(GLUT8) is a mammalian trehalose transporter required for trehalose-induced autophagy[J].Scientific Reports,2016,6:38586.
[23] CHANG H,LI X,CAI Q,et al.The PI3K/Akt/mTOR pathway is involved in CVB3-induced autophagy of HeLa cells[J].International Journal of Molecular Medicine,2017,40(1):182-192.
[24] MERRILL N M,SCHIPPER J L,KARNES J B,et al.PI3K-C2alpha knockdown decreases autophagy and maturation of endocytic vesicles[J].PLoS One,2017,12(9):e0184909.
[25] HUANG X,ZANG Y,ZHANG M,et al.Nuclear factor of κB1 is a key regulator for the transcriptional activation of milk synthesis in bovine mammary epithelial cells[J].DNA & Cell Biology,2017,36(4):295-302.
[26] OSORIO J S,LOHAKARE J,BIONAZ M.Biosynthesis of milk fat,protein,and lactose:Roles of transcriptional and post transcriptional regulation[J].Physiological Genomics,2016,48(4):231-256.
[27] KARIM M R,KADOWAKI M.Effect and proposed mechanism of vitamin C modulating amino acid regulation of autophagic proteolysis[J].Biochimie,2017,142:51-62.
[28] CHEN R,ZOU Y,MAO D,et al.The general amino acid control pathway regulates mTOR and autophagy during serum/glutamine starvation[J].The Journal of Cell Biology,2014,206(2):173-182.
[29] XIA X,CHE Y,GAO Y,et al.Arginine supplementation recovered the IFN-γ-mediated decrease in milk protein and fat synthesis by inhibiting the GCN2/eIF2α pathway,which induces autophagy in primary bovine mammary epithelial cells[J].Molecules & Cells,2016,39(5):410-417.
[30] FAN S J,GOBERDHAN D C I.PATs and SNATs:Amino acid sensors in disguise[J].Frontiers in Pharmacology,2018,9:640.
[31] SURYAWAN A,DAVIS T A.Regulation of protein synthesis by amino acids in muscle of neonates[J].Frontiers in Bioscience,2011,16:1445-1460.
[32] WANG C X,CHEN F,ZHANG W F,et al.Leucine promotes the growth of fetal pigs by increasing protein synthesis through the mTOR signaling pathway in longissimus dorsi muscle at late gestation[J].Journal of Agricultural and Food Chemistry,2018,66(15):3840-3849.

SNAT2: A Key Mediator for Methionine to Regulate Proliferation and Autophagy in Bovine Mammary Epithelial Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	XIAO Peng, SHANG Jianghua, YANG Chunyan, LI Mengqi, DUAN Anqin, MA Xiaoya, FENG Chao, HUANG Chenqian, ZHANG Bo, ZHOU Jinchen, WEI Kelong, ZHENG Wei, ZHENG Haiying. Effects of Alpha-linolenic Acid on Buffalo Ovarian Granulosa Cells Cultured in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2380-2387.
[2]	LI Can, LI Chuchu, ZENG Wei, ZHANG Ning, JI Chunxiao, CHEN Tao. Recombinant Expression and Function Study of RegⅢγ Protein in Porcine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2414-2426.
[3]	CHENG Qian, GAO Qingqing, WANG Yuhe, HUAN Changchao, GAO Song. Effects of Avian Pathogenic Escherichia coli Outer Membrane Protein OmpA on Autophagy of DF-1 Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1971-1980.
[4]	ZHEN Zhen, WANG Mei, WANG Yimin, HU Debao, ZHANG Linlin, LI Xin, GUO Yiwen, GUO Hong, DING Xiangbin. Effect of RNA Methylation Transfer Enzyme METTL3 on the Proliferation and Myogenic Differentiation of Bovine Skeletal Muscle Satellite Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1025-1036.
[5]	WEI Chunyan, GUO Jia, ZHU Dexin, ZHANG Wei, ZHU Jiale, DEN Xingmei, JIA Sifeng, LIU Liangbo, ZHANG Hui. Construction of Deletion Strain of Brucella BPE159 Gene and Effect of BPE159 Protein on Expression of Cellular Autophagy Factors [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 26-36.
[6]	TIAN Shuyue, SHEN Yingchao, WANG Xisheng, WANG Min, YI Minna, Tseweendolmaa, TAO Li, ZHAO Bilig, Manglai, Gerelchimeg. Study on Establishment of Immortalized Cell System of Equine Fetal Fibroblasts in Mongolian Horse [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 46-57.
[7]	TAN Haoyun, LIU Qian, HU Debao, ZHANG Linlin, LI Xin, DING Xiangbin, GUO Hong, GUO Yiwen. Effects of Interference lnc721 on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3292-3300.
[8]	WANG Xuefang, GUI Rong, CHEN Gongyi, WANG Wei, ZHANG Hongying, WANG Xuebing, LI Xiao. Clinical Study of Lianhuachaiqin Soluble Powder on Chicken Syndrome of Wind-heat Invading Lung [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3610-3621.
[9]	WANG Nan, FENG Baoliang, ZHENG Yunxi, HUANG Lei, WANG Yue, XU Songsong, ZHANG Xiuling, LIU Zhiguo, LI Kui, MU Yulian. Effects of WIP1 Gene on Proliferation and Differentiation of 3T3-L1 Preadipocytes and Its Expression in Different Growth Stages of Mice [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2869-2879.
[10]	ZHANG Xueping, SHI Bingang, JIN Xiayang, WANG Xiangyan, LAN Lijuan, SHI Yu, QI Youpeng, ZHAO Shijie, LI Shaobin, HU Jiang. Expression Characterization Analysis of KLF7 Gene and Effects of Its Overexpression on Proliferation and Differentiation of Preadipocytes in Tibetan Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2920-2930.
[11]	YU Kai, ZHAO Yufen, WENG Yu, WANG Qin, HAO Shaoyu, YU Boyang, DU Chenguang, SUBUDENG Gerile, LI Haijun. Effects of Dihydrotestosterone on Proliferation and Expression of Anti-Müllerian Hormone of Mouse Granulosa Cells [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3036-3043.
[12]	ZHOU Wei, LIU Xuping, ZHANG Yanmin, PENG Wenjuan, ZHAO Liang, TAN Wensong. Serum-free Suspension Culture of PK-15 Cells and Its Effect on the Proliferation of Porcine Circovirus Type 2 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2397-2405.
[13]	WANG Zihang, DENG Xingmei, QIU Runhui, ZHU Dexin, LI Jia, TAO Tingting, ZHU Jiale, SUN Zhihua, ZHANG Hui. Regulation of Long Noncoding RNAs in Autophagy Induced by PEDV Infected Vero-E6 Cells [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1942-1950.
[14]	LIU Fangjun, LAN Wutao, LI Pengfei, XU Dongmei, ZHANG Lei, ZHU Zhiwei. Effect of Tea Polyphenols on Proliferation and Migration of Mouse Melanoma Cell B16F10 in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1280-1286.
[15]	WANG Yueli, SHAO Zhiran, YI Jihai, WANG Yong, WANG Zhen, CHEN Chuangfu. miR-145a-3p Regulates Brucella Induced Mouse Macrophage RAW264.7 Autophagy by Targeting ATG14 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1117-1125.