Abstract
Long noncoding RNAs (lncRNAs) have been reported to play important roles in male reproduction. In our previous research, we studied the expression profile of lncRNAs in mouse male germ cells including spermatogonial stem cell, type A spermatogonia, pachytene spermatocyte, and round spermatid by microarray method, which showed that testis-enriched lncRNA AK015322 is highly expressed in spermatogonial stem cell. In this study, we found that AK015322 promotes proliferation of mouse spermatogonial stem cell line C18-4 in vitro. Furthermore, bioinformatic analysis, real-time PCR, and luciferase assay validated that AK015322 serves as a decoy of microRNA-19b-3p (miR-19b-3p), antagonizes its function, and attenuates the repression of its endogenous target transcriptional factor Ets-variant 5 (ETV5) which was a pivotal gene for spermatogonial stem cell self-renewal. Taken together, our results suggest that a variety of lncRNAs may regulate male reproduction through serving as competing-endogenous RNAs to modulate the function of germ cells.
Similar content being viewed by others
References
Agbor VA, Tao S, Lei N, Heckert LL (2013) A Wt1-Dmrt1 transgene restores DMRT1 to sertoli cells of Dmrt1(−/−) testes: a novel model of DMRT1-deficient germ cells. Biol Reprod 88(2):51
Akagi T, Kuure S, Uranishi K, Koide H, Costantini F, Yokota T (2015) ETS-related transcription factors ETV4 and ETV5 are involved in proliferation and induction of differentiation-associated genes in embryonic stem (ES) cells. J Biol Chem 290(37):22460–22473
Alankarage D, Lavery R, Svingen T, Kelly S, Ludbrook L, Bagheri-Fam S, Koopman P, Harley V (2016) SOX9 regulates expression of the male fertility gene Ets variant factor 5 (ETV5) during mammalian sex development. Int J Biochem Cell Biol
Anguera MC, Ma W, Clift D, Namekawa S, Kelleher RJ 3rd, Lee JT (2011) Tsx produces a long noncoding RNA and has general functions in the germline, stem cells, and brain. PLoS Genet 7(9):e1002248
Arun G, Akhade VS, Donakonda S, Rao MR (2012) Mrhl RNA, a long noncoding RNA, negatively regulates Wnt signaling through its protein partner Ddx5/p68 in mouse spermatogonial cells. Mol Cell Biol 32(15):3140–3152
Augoff K, McCue B, Plow EF, Sossey-Alaoui K (2012) miR-31 and its host gene lncRNA LOC554202 are regulated by promoter hypermethylation in triple-negative breast cancer. Mol Cancer 11:5
Bao J, Wu J, Schuster AS, Hennig GW, Yan W (2013) Expression profiling reveals developmentally regulated lncRNA repertoire in the mouse male germline. Biol Reprod 89(5):107
Cesana M, Cacchiarelli D, Legnini I, Santini T, Sthandier O, Chinappi M, Tramontano A, Bozzoni I (2011) A long noncoding RNA controls muscle differentiation by functioning as a competing endogenous RNA. Cell 147(2):358–369
Chen L, Wang W, Cao L, Li, Z, Wang X (2016) Long non-coding RNA CCAT1 acts as a competing endogenous RNA to regulate cell growth and differentiation in acute myeloid leukemia. Mol Cells 39(4):330–336
Chendrimada TP, Gregory RI, Kumaraswamy E, Norman J, Cooch N, Nishikura K, Shiekhattar R (2005) TRBP recruits the Dicer complex to Ago2 for microRNA processing and gene silencing. Nature 436(7051):740–744
Cui N, Hao G, Zhao Z, Wang F, Cao J, Yang A (2016) MicroRNA-224 regulates self-renewal of mouse spermatogonial stem cells via targeting DMRT1. J Cell Mol Med 20(8):1503–1512
Eo J, Shin H, Kwon S, Song H, Murphy KM, Lim JH (2011) Complex ovarian defects lead to infertility in Etv5−/− female mice. Mol Hum Reprod 17(9):568–576
Ganesan G, Rao SM (2008) A novel noncoding RNA processed by Drosha is restricted to nucleus in mouse. RNA 14(7):1399–1410
Golestaneh N, Beauchamp E, Fallen S, Kokkinaki M, Uren A, Dym M (2009) Wnt signaling promotes proliferation and stemness regulation of spermatogonial stem/progenitor cells. Reproduction 138(1):151–162
Ishii K, Kanatsu-Shinohara M, Toyokuni S, Shinohara T (2012) FGF2 mediates mouse spermatogonial stem cell self-renewal via upregulation of Etv5 and Bcl6b through MAP2K1 activation. Development 139(10):1734–1743
Johnston DS, Olivas E, DiCandeloro P, Wright WW (2011) Stage-specific changes in GDNF expression by rat Sertoli cells: a possible regulator of the replication and differentiation of stem spermatogonia. Biol Reprod 85(4):763–769
Kokkinaki M, Lee TL, He Z, Jiang J, Golestaneh N, Hofmann MC, Chan WY, Dym M (2009) The molecular signature of spermatogonial stem/progenitor cells in the 6-day-old mouse testis. Biol Reprod 80(4):707–717
Lee TL, Xiao A, Rennert OM (2012) Identification of novel long noncoding RNA transcripts in male germ cells. Methods Mol Biol 825:105–114
Leucci E, Patella F, Waage J, Holmstrom K, Lindow M, Porse B, Kauppinen S, Lund, AH (2013) microRNA-9 targets the long non-coding RNA MALAT1 for degradation in the nucleus. Sci Rep 3:2535
Lewis BP, Burge CB, Bartel DP (2005) Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120(1):15–20
Liang M, Li W, Tian H, Hu T, Wang L, Lin Y, Li Y, Huang H, Sun F (2014) Sequential expression of long noncoding RNA as mRNA gene expression in specific stages of mouse spermatogenesis. Sci Rep 4:5966
Liang WC, Fu WM, Wong CW, Wang Y, Wang WM, Hu GX, Zhang L, Xiao LJ, Wan DC, Zhang JF et al (2015) The lncRNA H19 promotes epithelial to mesenchymal transition by functioning as miRNA sponges in colorectal cancer. Oncotarget 6(26):22513–22525
Luk AC, Chan WY, Rennert OM, Lee TL (2014) Long noncoding RNAs in spermatogenesis: insights from recent high-throughput transcriptome studies. Reproduction 147(5):R131–R141
Monte D, Coutte L, Dewitte F, Defossez PA, Le Coniat M, Stehelin D, Berger R, de Launoit Y (1996) Genomic organization of the human ERM (ETV5) gene, a PEA3 group member of ETS transcription factors. Genomics 35(1):236–240
Morrow CM, Hostetler CE, Griswold MD, Hofmann MC, Murphy KM, Cooke PS, Hess RA (2007) ETV5 is required for continuous spermatogenesis in adult mice and may mediate blood testes barrier function and testicular immune privilege. Ann N Y Acad Sci 1120:144–151
Ni MJ, Hu ZH, Liu Q, Liu MF, Lu MH, Zhang JS, Zhang L, Zhang YL (2011) Identification and characterization of a novel non-coding RNA involved in sperm maturation. PLoS One 6(10):e26053
Niu Z, Goodyear SM, Rao S, Wu X, Tobias JW, Avarbock MR, Brinster RL (2011) MicroRNA-21 regulates the self-renewal of mouse spermatogonial stem cells. Proc Natl Acad Sci U S A 108(31):12740–12745
Oatley JM, Avarbock MR, Telaranta AI, Fearon DT, Brinster RL (2006) Identifying genes important for spermatogonial stem cell self-renewal and survival. Proc Natl Acad Sci U S A 103(25):9524–9529
Oatley JM, Avarbock MR, Brinster RL (2007) Glial cell line-derived neurotrophic factor regulation of genes essential for self-renewal of mouse spermatogonial stem cells is dependent on Src family kinase signaling. J Biol Chem 282(35):25842–25851
Romero Y, Meikar O, Papaioannou MD, Conne B, Grey C, Weier M, Pralong F, De Massy B, Kaessmann H, Vassalli JD et al (2011) Dicer1 depletion in male germ cells leads to infertility due to cumulative meiotic and spermiogenic defects. PLoS One 6(10):e25241
Salmena L, Poliseno L, Tay Y, Kats,L, Pandolfi PP (2011) A ceRNA hypothesis: the Rosetta Stone of a hidden RNA language? Cell 146(3):353–358
Song R, Ro S, Michaels JD, Park C, McCarrey JR, Yan W (2009) Many X-linked microRNAs escape meiotic sex chromosome inactivation. Nat Genet 41(4):488–493
Sun J, Lin Y, Wu J (2013) Long non-coding RNA expression profiling of mouse testis during postnatal development. PLoS One 8(10):e75750
Tay Y, Rinn J, Pandolfi PP (2014) The multilayered complexity of ceRNA crosstalk and competition. Nature 505(7483):344–352
Trupp M, Ryden M, Jornvall H, Funakoshi H, Timmusk T, Arenas E, Ibanez CF (1995) Peripheral expression and biological activities of GDNF, a new neurotrophic factor for avian and mammalian peripheral neurons. J Cell Biol 130(1):137–148
Tyagi G, Carnes K, Morrow C, Kostereva NV, Ekman GC, Meling DD, Hostetler C, Griswold M, Murphy KM, Hess RA et al (2009) Loss of Etv5 decreases proliferation and RET levels in neonatal mouse testicular germ cells and causes an abnormal first wave of spermatogenesis. Biol Reprod 81(2):258–266
Wang SH, Zhang WJ, Wu XC, Weng MZ, Zhang MD, Cai Q, Zhou D, Wang JD, Quan ZW (2016) The lncRNA MALAT1 functions as a competing endogenous RNA to regulate MCL-1 expression by sponging miR-363-3p in gallbladder cancer. J Cell Mol Med
Wu X, Goodyear SM, Tobias JW, Avarbock MR, Brinster RL (2011) Spermatogonial stem cell self-renewal requires ETV5-mediated downstream activation of Brachyury in mice. Biol Reprod 85(6):1114–1123
Wu C, Macleod I, Su AI (2013) BioGPS and MyGene.info: organizing online, gene-centric information. Nucleic Acids Res 41(Database issue):D561–D565
Yoshida S, Sukeno M, Nabeshima Y (2007) A vasculature-associated niche for undifferentiated spermatogonia in the mouse testis. Science 317(5845):1722–1726
Zhang Z, Cheng J, Wu Y, Qiu J, Sun Y, Tong X (2016) LncRNA HOTAIR controls the expression of Rab22a by sponging miR-373 in ovarian cancer. Mol Med Rep 14(3):2465–2472
Acknowledgments
We appreciate Dr. Fei Sun (Shanghai Jiaotong University, P.R. China) for the C18-4 and HEK293T cell lines. This study was supported by the Natural Science Foundation of Anhui Province (P.R. China) (1608085QH199), the Natural Science Foundation of Bengbu Medical College (P.R. China) (BYKY1406ZD, BYKY1444, and BYKY1421ZD), the Undergraduate Training Program for Innovation and Entrepreneurship of Anhui Province (P.R. China) (201510367044), and the Key Project of the Natural Science Foundation of Anhui Provincial Department of Education (P.R. China) (KJ2015A289).
Authors’ contribution
Ke Hu and Meng Liang conceived and performed this study; Meng Liang wrote the manuscript; and Jing Zhang reviewed the manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Editor: Tetsuji Okamoto
Rights and permissions
About this article
Cite this article
Hu, K., Zhang, J. & Liang, M. LncRNA AK015322 promotes proliferation of spermatogonial stem cell C18-4 by acting as a decoy for microRNA-19b-3p. In Vitro Cell.Dev.Biol.-Animal 53, 277–284 (2017). https://doi.org/10.1007/s11626-016-0102-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11626-016-0102-5