miR-23基因家族进化分析及其在猪睾丸组织中的表达变化
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摘要
运用生物信息学方法在miRBase数据库检索到miR-23基因家族的序列,利用Ensembl数据库信息确定miR-23在基因组的位置,采用MEGA5.0构建miR-23系统进化树,再利用RT-q PCR技术定量检测其在沙子岭猪睾丸组织中7个不同发育时期(D1、D30、D60、D90、D120、D150、D180)的表达变化。结果表明:在37种物种中共搜索到82条序列,且各序列具有高度保守性,大部分位于基因间隔区。进化分析表明:各物种miR-23基因家族可能以不同的方式和速度进化而来。RT-q PCR检测结果显示,miR-23基因家族在猪睾丸组织中不同发育时期表现出相同趋势的表达规律,即出生后其表达水平有所降低,到性成熟期又呈现升高趋势。本研究对miR-23基因家族进行系统进化分析并检测其在沙子岭猪睾丸组织中表达变化规律,为进一步探明miR-23基因家族的功能及作用机制奠定基础。
The sequences of miR-23 gene family were retrieved from the miRBase database using bioinformatics methods. The position of miR-23 in the genome was determined using the Ensembl database. The miR-23 phylogenetic tree was constructed using MEGA 5. 0,and then quantified using RT-qPCR. The expression of the gene in seven different developmental stages( at days 1,30,60,90,120,150 and 180) of the pig testis in Shaziling was detected. The results showed that 82 sequences were found in 37 species,and the sequences were highly conserved. Most of them were located in the intergenic region. Evolutionary analysis showed that the miR-23 gene family of various species might have evolved in different ways and at different speeds. The results of RT-qPCR showed that the miR-23 gene family showed the same trend of expression at different developmental stages in the pig testis tissues,which the expression level of the miR-23 gene family was decreased after birth and increased again during the sexual maturation period. In this study,the miR-23 gene family was systematically,phylogenetically analyzed and its expression patterns in the testis of pigs in Shaziling were examined,which laid the foundation for further exploration of the functions and mechanisms of the miR-23 gene family.
The sequences of miR-23 gene family were retrieved from the miRBase database using bioinformatics methods. The position of miR-23 in the genome was determined using the Ensembl database. The miR-23 phylogenetic tree was constructed using MEGA 5. 0,and then quantified using RT-qPCR. The expression of the gene in seven different developmental stages( at days 1,30,60,90,120,150 and 180) of the pig testis in Shaziling was detected. The results showed that 82 sequences were found in 37 species,and the sequences were highly conserved. Most of them were located in the intergenic region. Evolutionary analysis showed that the miR-23 gene family of various species might have evolved in different ways and at different speeds. The results of RT-qPCR showed that the miR-23 gene family showed the same trend of expression at different developmental stages in the pig testis tissues,which the expression level of the miR-23 gene family was decreased after birth and increased again during the sexual maturation period. In this study,the miR-23 gene family was systematically,phylogenetically analyzed and its expression patterns in the testis of pigs in Shaziling were examined,which laid the foundation for further exploration of the functions and mechanisms of the miR-23 gene family.
引文
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[3] Dongiovanni P,Romeo S,Valenti L.Hepatocellular carcinoma in nonalcoholic fatty liver:Role of environmental and genetic factors[J]. World Journal of Gastroenterology,2014,20(36):12945-12955.
[4] Ma Z L,Zhang B J,Wang D T,et al.Tanshinones suppress AURKA through up-regulation of miR-32 expression in non-small cell lung cancer[J].Oncotarget,2015,6(24):20111-20120.
[5] Shao Y,Shen Y Q,Li Y L,et al.Direct repression of the oncogene CDK4 by the tumor suppressor miR-486-5p in non-small cell lung cancer[J].Oncotarget,2016,7(23):34011.
[6] Zhang B,Farwell M A. microRNAs:a new emerging class of players for disease diagnostics and gene therapy[J]. Journal of Cellular&Molecular Medicine,2008,12(1):3-21.
[7]彭馥芝,冉茂良,翁波,等.miR-221/222基因家族进化分析及其在猪睾丸组织中的表达变化[J].经济动物学报,2017,21(1):33-40.
[8]翁波,彭馥芝,冉茂良,等.miR-24基因家族进化及其在猪睾丸组织中的表达变化分析[J].经济动物学报,2017,21(2):106-111.
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[10] Wang X,Wang J,Ma H,et al. Down regulation of miR-195correlates with lymph node metastasis and poor prognosis in colorectal cancer[J]. Medical Oncology,2012,29(2):919-927.
[11]罗丹丹,罗玉萍,彭娟,等.miRNA-9基因家族进化分析及其靶基因预测[J].中国细胞生物学学报,2011(5):510-519.
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[13]金科,朱剑梅,邱发麒.微小RNA-23a对乳腺癌细胞增殖和侵袭的影响[J].中国临床药理学杂志,2017,33(16):1568-1571.
[14]劳玲娟,宋新江,徐佳.miR-23a和ESRP1在直肠癌中的作用[J].中国病理生理杂志,2017,33(5):817-825.
[15]商京丽,孙树汉.MicroRNA-23a通过线粒体凋亡途径调节结肠癌化疗敏感性的机制研究[D].上海:第二军医大学,2011.
[16]周雪鹏.miR-23a通过靶向MTSS1促进结肠癌侵袭转移及临床意义[D].衡阳:南华大学,2011.
[17] Koller K,Das S,Leuschner I,et al.Identification of the transcription factor HOXB4 as a novel target of miR-23a[J].Genes Chromosomes&Cancer,2013,52(8):709-715.
[18] Viswanathan V,Fields J,Boman B M. The miRNA23bregulated signaling network as a key to cancer development—implications for translational research and therapeutics[J].Journal of Molecular Medicine-jmm,2014,92(11):1129-1138.
[19]徐立凤,禹小芳,张丽华,等.miR-23b-3p对3T3-L1前脂肪细胞增殖和成脂分化的影响[J].中国兽医学报,2017,37(10):2014-2019.
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[31] Yang X,Zhou Y,Peng S,et al.Differentially expressed plasma microRNAs in premature ovarian failure patients and the potential regulatory function of mir-23a in granulosa cell apoptosis[J].Reproduction,2012,144(2):235-244.
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