甘氨酰tRNA合成酶对奶牛乳腺上皮细胞增殖的影响

doi:10.16431/j.cnki.1671-7236.2016.06.024

›› 2016, Vol. 43 ›› Issue (6): 1557-1565.doi: 10.16431/j.cnki.1671-7236.2016.06.024

甘氨酰tRNA合成酶对奶牛乳腺上皮细胞增殖的影响

古新宇, 骆超超, 敖金霞, 黄鑫, 臧艳丽, 孙霞, 高学军

东北农业大学, 乳品科学教育部重点实验室, 哈尔滨 150030

收稿日期:2016-01-04 出版日期:2016-06-20 发布日期:2016-07-11
通讯作者: 高学军 E-mail:gaoxj5390@sina.com
作者简介:古新宇(1989-),女,黑龙江牡丹江人,硕士,研究方向:泌乳生物学与乳腺功能调控,E-mail:guxinyu33@163.com
基金资助:
科技部"863"项目(2013AA102504-03);国家自然科学基金项目(31472162)

Effect of Glycyl-tRNA Synthetase on Cell Proliferation of Bovine Mammary Epithelial Cells

GU Xin-yu, LUO Chao-chao, AO Jin-xia, HUANG Xin, ZANG Yan-li, SUN Xia, GAO Xue-jun

Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China

Received:2016-01-04 Online:2016-06-20 Published:2016-07-11

摘要/Abstract

摘要： 甘氨酰tRNA合成酶(glycyl-tRNA synthetase,GlyRS)是重要的氨基酰tRNA合成酶(aminoacyl-tRNA synthetases,aaRSs)家族成员。目前,国内外关于GlyRS 在奶牛乳腺发育过程中表达及调节的研究鲜有报道。为了揭示GlyRS的表达与奶牛乳腺发育与泌乳之间的关系,本试验应用EdU免疫荧光法、流式细胞术检测GlyRS对奶牛乳腺上皮细胞(bovine mammary epithelial cells,BMEC)中DNA数目及细胞周期的影响,实时荧光定量PCR、Western blotting技术检测GlyRS对BMEC中Cyclin D1的表达影响。结果显示,GlyRS过表达时,细胞的DNA合成、处于S期和G2/M期的细胞数、细胞中Cyclin D1的表达均增加;GlyRS抑制后,细胞的DNA合成、处于S期和G2/M期的细胞数、细胞中Cyclin D1的表达均减少。表明GlyRS通过上调Cyclin D1的表达,加快细胞周期转换,增加DNA合成数目,从而促进BMEC增殖。

关键词: 奶牛乳腺上皮细胞; 甘氨酰tRNA合成酶; 细胞增殖

Abstract: Glycyl-tRNA synthetase is an important member of aminoacyl-tRNA synthetase family.However,the knowledge about expression and regulation mechanism of GlyRS in the development of bovine mammary gland is limited.To reveal the relationship among GlyRS and bovine mammary gland development and lactation,EdU immunofluorescence and flow cytometry were used to analyze the impact of GlyRS on the number of DNA and the influence of the cell cycle of BMEC,and the expression of Cyclin D1 of BMEC by Real-time PCR and Western blotting.The results showed that the DNA synthesis of BMEC,the numbers of cells in S and G2/M phase,and Cyclin D1 expression levels in BMEC were all increased after BMEC was transfected with a GlyRS overexpression vector,whereas all of the above indexes were reduced after BMEC was transfected with a shRNA targeting against GlyRS.This study revealed that GlyRS accelerates cell cycle transformation,increases the number of DNA synthesis by upregulating the expression of Cyclin D1,thereby promoting BMEC proliferation.

Key words: bovine mammary epithelial cells; glycyl-tRNA synthetase; cell proliferation

中图分类号:

Q813.1

古新宇, 骆超超, 敖金霞, 黄鑫, 臧艳丽, 孙霞, 高学军. 甘氨酰tRNA合成酶对奶牛乳腺上皮细胞增殖的影响[J]. , 2016, 43(6): 1557-1565.

GU Xin-yu, LUO Chao-chao, AO Jin-xia, HUANG Xin, ZANG Yan-li, SUN Xia, GAO Xue-jun. Effect of Glycyl-tRNA Synthetase on Cell Proliferation of Bovine Mammary Epithelial Cells[J]. , 2016, 43(6): 1557-1565.

参考文献

[1] Hausmann C D,Ibba M.Aminoacyl-tRNA synthetase complexes:Molecular multitasking revealed[J].FEMS Microbiol Rev,2008,32(4):705-721.
[2] Nurit Y C,Irit C L,Kay G,et al.LysRS serves as a key signaling molecule in the immune response by regulating gene expression[J].Molecular Cell,2009,34:603-611.
[3] Xu X,Shi Y,Zhang H M,et al.Unique domain appended to vertebrate tRNA synthetase is essential for vascular development[J].Nat Commun,2012,3:681.
[4] Han J M,Jeong S J,Park M C,et al.Leucyl-tRNA synthetase is an intracellular leucine sensor for the mTORC1-signaling pathway[J].Cell,2012,149(2):410-424.
[5] Arif A,Jia J,Moodt R A,et al.Phosphorylation of glutamyl-prolyl tRNA synthetase by cyclin-dependent kinase 5 dictates transcript-selective translational control[J].Proc Natl Acad Sci USA,2011,108(4):1415-1420.
[6] 丁梅,刘小倩,高钓明,等.色氨酞tRNA合成酶对淋巴瘤小鼠细胞增殖的影响[J].山东大学学报,2011,49(12):48-52.
[7] Svitlana H,Marc M.Aminoacyl-tRNA synthetase complexes in evolution[J].Molecular Sciences,2015,16:6571-6594.
[8] 朱斌,王恩多.氨基酰-tRNA合成酶与神经退行性疾病[J].生物化学与生物物理进展,2007,34(6):562-566.
[9] Inoki K,Ouyang H,Zhu T,et al.TSC2 integrates Wnt and energy signals via a coordinated phosphorylation by AMPK and GSK3 to regulate cell growth[J].Cell,2006,126:955-968.
[10] Antonellis A,Ellsworth R E,Sambuughin N,et al.Glycyl-tRNA synthetase mutations in charcot-marie-tooth disease type 2D and distal spinal muscular atrophy type V[J].Am J Hum Genet,2003,72(5):1293-1299.
[11] Hugh J M,Jeremy S,Amanda S,et al.Compound heterozygous mutations in glycl-tRNA synthetase are a proposed cause of systemic mitochondrial disease[J].BMC Medical Genetics,2014,15:36-41.
[12] Jordanova A,Irobi J,Thomas F P,et al.Disrupted function and axonal distribution of mutant tyrosyl-tRNA synthetase in dominant intermediate charcot-marie-tooth neuropathy[J].Nature Genetics,2006,38:197-202.
[13] Latour P,Thauvin-Robinet C,Baudelet-Méry C,et al.A major determinant for binding and aminoacylation of tRNA(Ala) in cytoplasmic alanyl-tRNA synthetase is mutated in dominant axonal charcot-marie-tooth disease[J].American Journal of Human Genetics,2010,86:77-82.
[14] Vester A,Velez-Ruiz G,McLaughlin H M,et al.A loss-of-function variant in the human histidyl-tRNA synthetase(HARS) gene is neurotoxic in vivo[J].NIH Public Access,2013,34(1):191-199.
[15] Gonzalez M,McLaughlin H,Houlden H,et al.Exome sequencing identifies a significant variant in methionyl-tRNA synthetase(MARS) in a family with late-onset CMT2[J].NIH Public Access,2013,84(11):1247-1249.
[16] Mun J,Kim Y H,Yu J,et al.A proteomic approach based on multiple parallel separation for the unambiguous identification of an antibody cognate antigen[J].Electrophoresis,2010,31(20):3428-3436.
[17] 陆黎敏,李庆章,黄建国,等.双向电泳分析蛋氨酸对奶牛乳腺上皮细胞核磷酸化蛋白质的影响[J].中国畜牧兽医,2013,40(6):53-56.
[18] 黄建国,高学军,李庆章,等.双向电泳检测奶牛乳腺上皮细胞核磷酸化蛋白质方法的建立[J].乳业科学与技术,2011,34(6):250-253.
[19] 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):27-31.
[20] Hu H,Wang J,Bu D,et al.In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow[J].PLoS One,2009,4(11):e7636.
[21] Livak K J,Schmittgen T D.Analysis of relative gene expression data using Real-time quantitative PCR and the 2(-delta delta C(T)) method[J].Methods,2001,25(4):402-408.
[22] 王卓然,侯晓明,曲波,等.Pten对奶牛乳腺上皮细胞活力及乳蛋白合成的影响[J].中国生物化学与分子生物学报,2014,30(4):395-401.
[23] Boutinaud M,Guinard-Flamenta J,Jammes H,et al.The number and activity of mammary epithelial cells,determining factors for milk production[J].Reprod Nutr Dev,2004,44(5):499-508.
[24] Zeng C,Pan F,Jones L A,et al.Evaluation of 5-ethynyl-2'-deoxyuridine staining as a sensitive and reliable method for studying cell proliferation in the adult nervous system[J].Brain Res,2010,1319:21-32.
[25] 张霞,高学军,赵锋,等.糖原合成酶激酶3β对奶牛乳腺上皮细胞增殖及细胞周期的影响[J].中国畜牧兽医,2014,41(4):128-132.

甘氨酰tRNA合成酶对奶牛乳腺上皮细胞增殖的影响

Effect of Glycyl-tRNA Synthetase on Cell Proliferation of Bovine Mammary Epithelial Cells

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