Abstract:MicroRNAs are a class of tiny, single-stranded noncoding RNAs that can target mRNA via base pairing and lead to translational repression or mRNA degradation at the level of gene posttranscription, which are involved in organ development, cell differentiation, proliferation, apoptosis and other biological processes, closely related with many diseases. Basic leucine zipper and W2 domains 2 (BZW2) functions in protein synthesis metabolism. The purpose of this study was to validate whether miR-122 could inhibit BZW2 gene expression in chicken (Gallus gallus). The bioinformatics method was used to investigate whether miR-122 could pair chicken BZW2 3'-UTR, furthermore dual-luciferase report system and mutant assay were used to validate that miR-122 repressed BZW2 expression. qRT-PCR was used to detect the expression level of BZW2 in leghorn hepatocellar (LHM) cells transfected with miRNA-122mimic and primary chicken hepatocytes transfected with LNA-antimiR-122. Bioinformatics prediction and dual luciferase report system proved that BZW2 was target gene of miR-122, target site mutation assay showed the complementary site of 3'UTR with miR-122 seeds region were target site of miR-122. After overexpression of miR-122 in leghorn hepatocellar (LMH) cells, BZW2 mRNA expression level decreased obviously. Using LNA-antimiR-122 to knockdown the miR-122 expression in primary chicken hepatocytes, the target gene BZW2 mRNA expression level increased significantly. These data suggested that BZW2 gene was target gene of miR-122 and BZW2 mRNA was negatively regulated by miR-122 in chicken hepatocytes. It will provide scientific basis on revealing a wide range of functions in the liver of miR-122 in chicken.
[1]Bartel D P.MicroRNAs: genomics,biogenesis,mechanism,and function[J].Cell, 2004, 116(2):281-297
[2]Lee R C, Feinbaum R L, Ambros V.The Celegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14[J].Cell, 1993, 75(5):843-854
[3]Doench J G, Sharp P A.Specificity of microRNA target selection in translational repression[J].Genes & Develoment, 2004, 18(5):504-511
[4]Lagos-Quintana M, Rauhut R, Yalcin A, Meyer J, Lendeckel W, Tuschl T.Identification of tissue-specific microRNAs from mouse[J].Current Biology, 2002, 12(9):735-739
[5]Chang J, Nicolas E, Marks D, et al.miR-122,a mammalian liver-specific microRNA,is processed from hcr mRNA and may downregulate the high affinity cationic amino acid transporter CAT-I[J].RNA Biology, 2004, 1(2):106-13
[6]Sun R, Li Q, Yan H, et al.Bzw2 Promotes Proliferation and Lactation of Mammary Epithelial Cell in Dairy Goat[J].Journal of Integrative Agriculture, 2012, 11(11):1884-91
[7] Wei Zhi-yi.Structural Biology Study of Eukaryotic translation initiation factor eIF3 and eIF5[D]. University of China Science and Technology, 2007.
[8]魏志毅.真核翻译起始因子eIF3和eIF5的结构生物学研究 [D]. 中国科学科技大学, 2007
[9]Asano K, Clayton J, Shalev A, et al.A multifactor complex of eukaryotic initiation factors eIF1,eIF2,eIF3,eIF5,and initiator tRNAMet is an important translation initiation intermediate in vivo[J].Genes & Development, 2000, 14(19):2534-46
[10]Singh CR, Watanabe R, Zhou D, et al.Mechanisms of translational regulation by a human eIF5-mimic protein[J].Nucleic Acids Research, 2011, 39(19):8314-28
[11]Wang Xingguo, Shao Fang, Yu Jianfeng, Jiang Honglin, Gong Daoqing, Gu Zhiliang.MicroRNA-122 targets genes related to liver metabolism in chickens[J].[J].Comparative Biochemistry and Physiology( Part B),, 2015, , 184(1):29-35
[12]Wang Hui-juan, Yang Lei, Shao Fang, Wang Xing-guo, Yu Jian-feng, Gu Zhi-liang.Research on GH Regulation of GHIGF-Ⅰ Pathway Related Gene Expression in Primary Chicken Hepatocytes[J].Journal of Yangzhou University(Agricultural and Life Science Edition), 2013, 34(3):23-28
[13]王慧娟,杨磊,邵芳,王星果,郁建锋,顾志良,.GH调控鸡原代肝细胞中GH/IGF-Ⅰ信号通路相关基因mRNA表达的研究[J].扬州大学学报(农业和生命科学版), 2013, 34(3):23-28
[14] Shao Fang, Wang Xingguo, Yu Jianfeng, Jiang Honglin, Zhu Bin, Gu Zhiliang.Expression of miR-33 from anSREBF2 Intron Targets the FTO Gene in the Chicken[J]. [J].PLOS One, 2014, 9(e91236):1-8
[15]Enright AJ, John B, Gaul U, et al.MicroRNA targets in Drosophila[J].Genome Biology, 2003, 5(1):R1-6
[16]Lewis BP, Burge CB, Bartel DP.Conserved seed pairing,often flanked by adenosines,indicates that thousands of human genes are microRNA targets[J].Cell, 2005, 120(1):15-20
[17]Krek A, Grun D, Poy MN, et al.Combinatorial microRNA target predictions[J].Nature Genetics, 2005, 37(5):495-500
[18]Li J, Ghazwani M, Zhang Y, et al.miR-122 regulates collagen production via targeting hepatic stellate cells and suppressing P4HA1 expression[J].Journal of Hepatology, 2013, 58(3):522-8
[19]Boutz DR, Collins PJ, Suresh U, et al.Two-tiered approach identifies a network of cancer and liver disease-related genes regulated by miR-122[J].Journal of Biological Chemistry, 2011, 286(20):18066-78
[20]Jung CJ, Iyengar S, Blahnik KR, et al.Epigenetic modulation of miR-122 facilitates human embryonic stem cell self-renewal and hepatocellular carcinoma proliferation[J].PLoS One, 2011, 6(11):e27740-8
[21]Karakatsanis A, Papaconstantinou I, Gazouli M, et al.Expression of miRNAs,miR-21,miR-31,miR-122,miR-145,miR-146a,miR-200c,miR-221,miR-222,and miR-223 in patients with hepatocellular carcinoma or intrahepatic cholangiocarcinoma and its prognostic significance[J].Molecular Carcinogenesis, 2013, 52(4):297-303
[22] Zhuang Peng,Li Zhi-Ying,Wang Xiang-Chen,Zheng Chang-Jing.The functional characterization of the human liver-specific miR-122 expression in hepatocullar carcinoma [J][J].Chinese Journal of Immunology, 2013, 29(7):686-688
[23]庄鹏, 李志莹, 王湘郴.肝特异性的 -对肝癌细胞增殖及凋亡的影响[J].中国免疫学杂志, 2013, 29(7):686-8