牦牛miR-378前体克隆及组织表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0671

生物技术通报 ›› 2019, Vol. 35 ›› Issue (1): 58-66.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0671

牦牛miR-378前体克隆及组织表达分析

纪会¹, 王会¹, 柴志欣¹, 王吉坤¹, 罗晓林², 姬秋梅³, 信金伟³, 钟金城¹

1. 西南民族大学青藏高原动物遗传资源保护与利用教育部重点实验室,成都 610041;
2. 四川省草原科学研究院,成都 611731;
3. 西藏自治区农牧科学院省部共建青稞和牦牛种质资源与遗传改良国家重点实验室,拉萨 850009

收稿日期:2018-07-23 出版日期:2019-01-26 发布日期:2019-01-23
作者简介:纪会,女,硕士研究生,研究方向：动物遗传学;E-mail：1328051168@qq.com
基金资助:
西南民族大学研究生创新型科研项目（CX2018SZ113）,国家肉牛牦牛产业技术体系项目（CARS-37）

Precursor Cloning and Tissue Expression Analysis of Yak miR-378

JI Hui¹, WANG Hui¹, CHAI Zhi-xin¹, WANG Ji-kun¹, LUO Xiao-lin², JI Qiu-mei³, XIN Jin-wei³, ZHONG Jin-cheng¹

1. Key Laboratory of Animal Genetics and Breeding of the Ministry of Education,Southwest Minzu University,,Chengdu 610041;
2. Sichuan Grassland Science Academy,Chengdu 611731;
3. State Key Laboratory of Barley and Yak Germplasm Resources and Genetic Improvement,Tibet Academy of Agricultural and Animal Husbandry Sciences,Lhasa 850009

Received:2018-07-23 Published:2019-01-26 Online:2019-01-23

摘要/Abstract

摘要： 旨在克隆牦牛miR-378的前体序列,阐明其组织表达规律,结合bta-miR-378靶基因的生物信息学预测和分析,探讨miR-378在牦牛生长发育过程中的调控功能。采用PCR方法成功克隆类乌齐牦牛miR-378前体序列,实时荧光定量PCR（RT-qPCR）检测miR-378-3p在各组织中的表达模式,结合生物信息学软件TargetScan、DAVID以及数据库NCBI、miRbase等对miR-378进行保守性分析、靶基因预测及其生物学功能分析。结果表明,miR-378在各物种间高度保守,且miR-378-3p在各组织中广泛表达,其中在臀大肌中表达水平最高,显著高于其他组织（P<0.01）,在臀脂中的表达高于卵巢、大脑、乳腺和肝脏。获得的272个靶基因主要参与细胞分化、细胞发育、大分子代谢等多个生物学过程,涉及孕酮介导的卵母细胞成熟、促性腺激素释放激素（GnRH）信号通路等,由此推测,miR-378可能在卵泡发育、卵母细胞成熟过程中起关键作用,进而影响母牦牛的繁殖性能。

关键词: 牦牛, miR-378, 克隆, 组织表达, 生物信息分析

Abstract: This work aims to clone the precursor sequence of yak miR-378,to elucidate its tissue expression pattern,and to explore the regulatory function of miR-378 in the growth and development of yak combined with bioinformatics prediction and analysis of bta-miR-378 target gene. PCR was employed to successfully clone the miR-378 precursor sequence of leiwuqi yak,and real-time quantitative PCR（RT-qPCR）was used to detect the expression patterns of miR-378-3p in various tissues. Further,bioinformatics software and databases such as TargetScan,DAVID,NCBI,miRbase,etc were together applied to perform conservative analysis,target gene prediction and biological function analysis of miR-378. The results showed that miR-378 was highly conserved among different species and miR-378-3p was widely expressed in all tissues,the expression level was the highest in gluteus maximus,which was significantly higher than that in other tissues（P < 0.01）. Expression in hip fat was higher than in ovary,brain,breast and liver. The obtained 272 target genes were mainly involved in many biological processes such as cell differentiation,cell development,and macromolecular metabolism,involved in progesterone-mediated oocyte maturation and gonadotropin releasing hormone（GnRH）signaling pathways. It is speculated that miR-378 may play a key role in follicular development and oocyte maturation,which may affect the reproductive performance of female calves.

Key words: yak, miR-378, cloning, tissue expression, biological information analysis

纪会, 王会, 柴志欣, 王吉坤, 罗晓林, 姬秋梅, 信金伟, 钟金城. 牦牛miR-378前体克隆及组织表达分析[J]. 生物技术通报, 2019, 35(1): 58-66.

JI Hui, WANG Hui, CHAI Zhi-xin, WANG Ji-kun, LUO Xiao-lin, JI Qiu-mei, XIN Jin-wei, ZHONG Jin-cheng. Precursor Cloning and Tissue Expression Analysis of Yak miR-378[J]. Biotechnology Bulletin, 2019, 35(1): 58-66.

参考文献

[1] Xie Z, Kasschau KD, Carrington AJC.Negative feedback regulation of in by microRNA-Guided mRNA degradation[J]. Current Biology, 2003, 13(9):784-789.
[2] Thermann R, Hentze MW.Drosophila miR2 induces pseudo-polysomes and inhibits translation initiation[J]. Nature, 2012, 447(7146):875-878.
[3] Lee RC, Feinbaum RL, Ambros V.The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14[J]. Cell, 1993, 75(5):843.
[4] Tong HL, Jiang RY, Liu TT, et al.bta-miR-378 promote the differentiation of bovine skeletal muscle-derived satellite cells[J]. Gene, 2018, 668:246-251.
[5] Kasashima K, Nakamura Y, Kozu T.Altered expression profiles of microRNAs during TPA-induced differentiation of HL-60 cells[J]. Biochem Biophys Res Commun, 2004, 322(2):403-410.
[6] Chen QG, Zhou W, Han T, et al.MiR-378 suppresses prostate cancer cell growth through downregulation of MAPK1 in vitro and in vivo[J]. Tumour Biol, 2016, 37(2):1-9.
[7] Zeng M, Zhu L, Li L, et al.miR-378 suppresses the proliferation, migration and invasion of colon cancer cells by inhibiting SDAD1[J]. Cellular & Molecular Biology Letters, 2017, 22(1):12.
[8] Fu H, Zhang J, Pan T, et al.miR-378a enhances the sensitivity of liver cancer to sorafenib by targeting VEGFR, PDGFRβ and c-Raf[J]. Molecular Medicine Reports, 2018, 17(3):4581-4588.
[9] Ji KX, Cui F, Qu D, et al.MiR-378 promotes the cell proliferation of non-small cell lung cancer by inhibiting FOXG1[J]. European Review for Medical & Pharmacological Sciences, 2018, 22(4):1011.
[10] Li S, Yang F, Wang M, et al.miR-378 functions as an onco-miRNA by targeting the ST7L/Wnt/β-catenin pathway in cervical cancer[J]. International Journal of Molecular Medicine, 2017, 40(4):1047.
[11] Chai SH, Noor SM, Nagoor NH.MiR-378 and MiR-1827 regulate tumor invasion, migration and angiogenesis in human Lung adenocarcinoma by Targeting RBX1 and CRKL, respectively[J]. Journal of Cancer, 2018, 9(2):331-345.
[12] Tan D, Zhou C, Han S, et al.MicroRNA-378 enhances migration and invasion in cervical cancer by directly targeting autophagy-related protein 12[J]. Molecular Medicine Reports, 2018, 17(5):6319-6326.
[13] Chan JK, Kiet TK, Blansit K, et al.MiR-378 as a biomarker for response to anti-angiogenic treatment in ovarian cancer[J]. Gynecologic Oncology, 2014, 133(3):568-574.
[14] Ma TH, Cheng L, Wang B, et al.MIR-378 induces apoptsis of granulosa cells during follicle development in cattle[J]. Journal of Animal & Plant Sciences, 2017, 27(5):1738-1742.
[15] Lu Z, Du L, Liu R, et al.MiR-378 and BMP-Smad can influence the proliferation of sheep myoblast[J]. Gene, 2018, 674:143-150.
[16] 张阳阳. miR-378在牛前体脂肪细胞分化的作用与机制[D]. 长春:吉林大学, 2014.
[17] 茹松伟, 申卫红, 杨鹏程, 等. microRNA靶基因预测算法研究概况及发展趋势[J]. 生命科学, 2007, 19(5):562-567.
[18] Yong Z, Li C, Hu L, et al.miR-378 activates the pyruvate-PEP futile cycle and enhances lipolysis to ameliorate obesity in mice[J]. Ebiomedicine, 2016, 5:93-104.
[19] Wei X, Hui L, Zhang B, et al.miR-378a-3p promotes differentiation and inhibits proliferation of myoblasts by targeting HDAC4 in skeletal muscle development[J]. RNA Biology, 2016, 13(12):1300-1309.
[20] Nunu H, Jiu W, Weidong X, et al.MiR-378a-3p enhances adipogenesis by targeting mitogen-activated protein kinase 1[J]. Biochemical & Biophysical Research Communications, 2015, 457(1):37-42.
[21] Liu SY, Zhang YY, Gao Y, et al.MiR-378 plays an important role in the differentiation of bovine preadipocytes[J]. Cellular Phy-siology & Biochemistry, 2015, 36(4):1552-1562.
[22] Xu S, Linhermelville K, Yang BB, et al.Micro-RNA378(miR-378)regulates ovarian estradiol production by targeting aromatase[J]. Endocrinology, 2011, 152(10):3941-3951.
[23] Toms D, Xu S, Pan B, et al.Progesterone receptor expression in granulosa cells is suppressed by microRNA-378-3p[J]. Molecular & Cellular Endocrinology, 2015, 399:95-102.
[24] Ma T, Jiang H, Gao Y, et al.Microarray analysis of differentially expressed microRNAs in non-regressed and regressed bovine corpus Luteum tissue;microRNA-378 may suppress Luteal cell apoptosis by targeting the interferon gamma receptor 1 gene[J]. Journal of Applied Genetics, 2011, 52(4):481-486.
[25] 徐盛玉. 营养水平对后备母猪卵巢microRNA-378表达影响及microRNA-378作用机制研究[D]. 成都:四川农业大学, 2012.
[26] 熊显荣, 兰道亮, 李键, 等. 牦牛卵巢小RNA高通量测序及生物信息学分析[J]. 畜牧兽医学报, 2016, 47(1):55-63.
[27] 潘阳阳, 崔燕, 樊江峰, 等. 胰岛素样生长因子-1(IGF-1)对牦牛卵丘细胞热休克蛋白70(HSP70)表达的影响及其与细胞凋亡的关联性分析[J]. 农业生物技术学报, 2015, 23(9):1207-1216.
[28] 李莉华, 高琪, 王晓艳, 等. miR-378靶向胰岛素样生长因子1类受体抑制肝癌细胞生长[J]. 中华肝脏病杂志, 2013, 21(8):609-613.

牦牛miR-378前体克隆及组织表达分析

Precursor Cloning and Tissue Expression Analysis of Yak miR-378

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