鸡miR-133a靶向调控BIRC5基因的表达

doi:10.3864/j.issn.0578-1752.2013.07.015

摘要/Abstract

摘要： 【目的】研究miR-133a在鸡不同组织中的表达情况，对baculoviral IAP repeat containing 5（BIRC5）进行靶基因的试验验证，探讨鸡miR-133a对BIRC5的靶向调控作用。【方法】采用生物信息学方法对miR-133a靶基因进行预测，并用双荧光素酶报告系统及点突变试验对BIRC5进行靶基因验证，同时运用实时荧光定量PCR检测miR-133a在鸡不同组织中的表达量。【结果】在鸡3′UTR数据库的11 891个基因中预测到287个miR-133a的靶基因；miR-133a在鸡的组织表达谱中显示其在肌肉中的表达量较高，尤其是在骨骼肌中表达量最高；报告基因试验显示BIRC5是miR-133a的靶基因，点突变试验证实了miR-133a在BIRC5上结合的靶位点。【结论】miR-133a是与鸡骨骼肌发育高度相关的miRNA，且鸡BIRC5基因是miR-133a的靶基因。

关键词: 鸡 , 骨骼肌 , miR-133a , BIRC5基因

Abstract: 【Objective】This study was conducted to investigate the expression pattern of miR-133a in various chicken tissues, and find whether baculoviral IAP repeat containing 5 (BIRC5) is the target gene of miR-133a.【Method】The bioinformatics methods were used to predict the target genes of miR-133a. Real-time PCR was used to detect the expression patterns of miR-133a of various tissues in chicken. Dual-luciferase reporter assays and site mutation assays were used to verify whether BIRC5 is a target gene of miR-133a. 【Result】Two hundred and eighty-seven genes of 11891 genes from 3′UTR database were predicted as target genes of miR-133a. The expression patterns of miR-133a in various chicken tissues showed that miR-133a was highly expressed in muscle especially in skeletal muscle. Reporter assays showed that BIRC5 was the target gene of miR-133a and the site mutation assays validated the target site of miR-133a in BIRC5. 【Conclusion】 miR-133a is a miRNA related with skeletal muscle development of chicken, and BIRC5 is a bona fide target gene in chicken.

Key words: chicken , skeletal muscle , miR-133a , BIRC5 gene

王星果, 邵芳, 龚道清, 卢祥云, 顾志良. 鸡miR-133a靶向调控BIRC5基因的表达[J]. 中国农业科学, 2013, 46(7): 1441-1447.

WANG Xing-Guo, SHAO Fang, GONG Dao-Qing, LU Xiang-Yun, GU Zhi-Liang. miR-133a Targets BIRC5 to Regulate Its Gene Expression in Chicken[J]. Scientia Agricultura Sinica, 2013, 46(7): 1441-1447.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2013.07.015

https://www.chinaagrisci.com/CN/Y2013/V46/I7/1441

参考文献

[1]Bartel D P. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell, 2004, 116(2): 281-297.

[2]郑永霞, 焦炳华. miRNA的生物形成及调控基因表达机制. 生命的化学, 2010, 30(6): 821-826.

Zheng Y X, Jiao B H. Biogenesis of microRNA and mechanism of the regulation of gene expression. Chemistry of Life, 2010, 30(6): 821-826. (in Chinese)

[3]王星果, 郁建锋, 顾志良. microRNA在肌肉发育中的功能研究进展. 生命科学, 2010, 22(2): 133-138.

Wang X G, Yu J F, Gu Z L. Function of microRNA in muscle development. Chinese Bulletin of Life Sciences, 2010, 22(2): 133-138. (in Chinese)

[4]McCarthy J J, Esser K A. MicroRNA-1 and microRNA-133a expression are decreased during skeletal muscle hypertrophy. Journal of Applied Physiology, 2007, 102(1): 306-313.

[5]Chen J F, Mandel E M, Thomson J M, Wu Q, Callis T E, Hammond S M, Conlon F L, Wang D Z. The role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation. Nature Genetics, 2006, 38(2): 228-233.

[6]Huang M B, Xu H, Xie S J, Zhou H, Qu L H. Insulin-like growth factor-1 receptor is regulated by microRNA-133 during skeletal myogenesis. PLoS One, 2011, 6(12): e29173.

[7]Care A, Catalucci D, Felicetti F, Bonci D, Addario A, Gallo P, Bang M L, Segnalini P, Gu Y, Dalton N D, Elia L, Latronico M V, Hoydal M, Autore C, Russo M A, Dorn G W, Ellingsen O, Ruiz-Lozano P, Peterson K L, Croce C M, Peschle C, Condorelli G. MicroRNA-133 controls cardiac hypertrophy. Nature Medicine, 2007, 13(5): 613-618.

[8]Belevych A E, Sansom S E, Terentyeva R, Ho H T, Nishijima Y, Martin M M, Jindal H K, Rochira J A, Kunitomo Y, Abdellatif M, Carnes C A, Elton T S, Gyorke S, Terentyev D. MicroRNA-1 and -133 increase arrhythmogenesis in heart failure by dissociating phosphatase activity from RyR2 complex. PLoS One, 2011, 6(12): e28324.

[9]Tao J, Wu D, Xu B, Qian W, Li P, Lu Q, Yin C, Zhang W. microRNA-133 inhibits cell proliferation, migration and invasion in prostate cancer cells by targeting the epidermal growth factor receptor. Oncology Reports, 2012, 27(6): 1967-1975.

[10]项茹, 吴正升, 张晴, 徐晓春, 吴强. miR-133a在乳腺癌组织中的表达及其临床意义. 安徽医科大学学报, 2011, 46(2): 109-112.

Xiang R, Wu Z S, Zhang Q, Xu X C, Wu Q. Expression of miR-133a in breast carcinoma and its significance. Acta Universitatis Medicinalis Anhui, 2011, 46(2): 109-112. (in Chinese)

[11]谢海涛. miR-133a在结肠癌组织中的表达. 广东医学, 2012, 33(15): 2242-2243.

Xie H T. Expression of miR-133a in colon carcinoma. Guangdong Medical Journal, 2012, 33(15): 2242-2243. (in Chinese)

[12]Ambrosini G, Adida C, Altieri D C. A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nature Medicine, 1997, 3(8): 917-921.

[13]Dallaglio K, Marconi A, Pincelli C. Survivin: a dual player in healthy and diseased skin. The Journal of Investigative Dermatology, 2012, 132(1): 18-27.

[14]Li F, Ambrosini G, Chu E Y, Plescia J, Tognin S, Marchisio P C, Altieri D C. Control of apoptosis and mitotic spindle checkpoint by survivin. Nature, 1998, 396(6711): 580-584.

[15]Li F, Ackermann E J, Bennett C F, Rothermel A L, Plescia J, Tognin S, Villa A, Marchisio P C, Altieri D C. Pleiotropic cell-division defects and apoptosis induced by interference with survivin function. Nature Cell Biology, 1999, 1(8): 461-466.

[16]Barrett R M, Colnaghi R, Wheatley S P. Threonine 48 in the BIR domain of survivin is critical to its mitotic and anti-apoptotic activities and can be phosphorylated by CK2 in vitro. Cell Cycle, 2011, 10(3): 538-548.

[17]Wang X G, Yu J F, Zhang Y, Gong D Q, Gu Z L. Identification and characterization of microRNA from chicken adipose tissue and skeletal muscle. Poultry Science, 2012, 91(1): 139-149.

[18]Li T, Wu R, Zhang Y, Zhu D. A systematic analysis of the skeletal muscle miRNA transcriptome of chicken varieties with divergent skeletal muscle growth identifies novel miRNAs and differentially expressed miRNAs. BMC Genomics, 2011, 12: 186.

[19]Lewis B P, Shih I H, Jones-Rhoades M W, Bartel D P, Burge C B. Prediction of mammalian microRNA targets. Cell, 2003, 115(7): 787-798.

[20]Lewis B P, Burge C B, Bartel D P. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell, 2005, 120(1): 15-20.

[21]Chen C, Ridzon D A, Broomer A J, Zhou Z, Lee D H, Nguyen J T, Barbisin M, Xu N L, Mahuvakar V R, Andersen M R, Lao K Q, Livak K J, Guegler K J. Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Research, 2005, 33(20): e179.

[22]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. Methods, 2001, 25(4): 402-408.

[23]Nicoli S, Standley C, Walker P, Hurlstone A, Fogarty K E, Lawson N D. MicroRNA-mediated integration of haemodynamics and Vegf signalling during angiogenesis. Nature, 2010, 464(7292): 1196-1200.

[24]Zhao Y, Samal E, Srivastava D. Serum response factor regulates a muscle-specific microRNA that targets Hand2 during cardiogenesis. Nature, 2005, 436(7048): 214-220.

[25]Zhao Y, Ransom J F, Li A, Vedantham V, von Drehle M, Muth A N, Tsuchihashi T, McManus M T, Schwartz R J, Srivastava D. Dysregulation of cardiogenesis, cardiac conduction, and cell cycle in mice lacking miRNA-1-2. Cell, 2007, 129(2): 303-317.

[26]Naguibneva I, Ameyar-Zazoua M, Polesskaya A, Ait-Si-Ali S, Groisman R, Souidi M, Cuvellier S, Harel-Bellan A. The microRNA miR-181 targets the homeobox protein Hox-A11 during mammalian myoblast differentiation. Nature Cell Biology, 2006, 8(3): 278-284.

[27]Stern-Ginossar N, Gur C, Biton M, Horwitz E, Elboim M, Stanietsky N, Mandelboim M, Mandelboim O. Human microRNAs regulate stress-induced immune responses mediated by the receptor NKG2D. Nature Immunology, 2008, 9(9): 1065-1073.