Please wait a minute...
Journal of Integrative Agriculture  2019, Vol. 18 Issue (8): 1924-1935    DOI: 10.1016/S2095-3119(19)62564-5
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene  
WENG Bo*, RAN Mao-liang*, Cao Rong, PENG Fu-zhi, LUO Hui, GAO Hu, TANG Xiang-wei, Yang An-qi, CHEN Bin
College of Animal Science and Technology/Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Changsha 410128, P.R.China
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
MicroRNAs (miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells.  However, the function roles of most of the miRNAs that have been identified in Sertoli cells are poorly understood.  In the present study, six experiments were conducted to study the regulatory role of miR-10b in porcine immature Sertoli cells.  In experiment 1, the results showed that the relative mRNA expression level of miR-10b in porcine testicular tissues decreased quadratically (P<0.001) with increasing age, while the relative mRNA expression level of DAZAP1 gene increased (P<0.001).  In addition, the mRNA expression of miR-10b was negatively (P<0.01) correlated with DAZAP1 mRNA expression (r=–0.550).  In experiment 2, the results from the bioinformatic analysis and a luciferase reporter assay demonstrated that miR-10b directly targeted the DAZAP1 gene in porcine immature Sertoli cells.  DAZAP1 mRNA and protein expressions were both regulated (P<0.05) by miR-10b.  In experiments 3 to 5, the over-expression of miR-10b or the siRNA-mediated knockdown of the DAZAP1 gene promoted (P<0.05) porcine immature Sertoli cell proliferation, as determined by the Cell Counting Kit-8 (CCK-8) assay and the 5-Ethynyl-2´-deoxyuridine (EdU) assay.  However, an annexin V-FITC/PI staining assay and the expression of cell survival-related genes indicated that over-expression of miR-10b or knockdown of DAZAP1 had no effect (P>0.05) on porcine immature Sertoli cell apoptosis.  In experiment 6, the co-transfection treatment results showed that miR-10b promoted (P<0.05) porcine immature Sertoli cell proliferation by targeting DAZAP1 gene.  Overall, these experiments demonstrated that miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene.
Keywords:  miR-10b        DAZAP1        gene expression        proliferation        porcine immature Sertoli cell  
Received: 28 May 2018   Accepted: 02 August 2019
Fund: This work was financially supported by the earmarked fund for China Agriculture Research System (CARS-36) and the Hunan Provincial Natural Science Foundation of China (2018JJ2176 and 2018JJ3219).
Corresponding Authors:  Correspondence CHEN Bin, Tel: +86-731-84618176, E-mail:    
About author:  WENG Bo, Tel: +86-731-84635385, E-mail:; * These authors contributed equally to this study.

Cite this article: 

WENG Bo, RAN Mao-liang, Cao Rong, PENG Fu-zhi, LUO Hui, GAO Hu, TANG Xiang-wei, Yang An-qi, CHEN Bin. 2019.

miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene  
. Journal of Integrative Agriculture, 18(8): 1924-1935.

Abu-Halima M, Backes C, Leidinger P, Keller A, Lubbad A M, Hammadeh M, Meese E. 2014. MicroRNA expression profiles in human testicular tissues of infertile men with different histopathologic patterns. Fertility and Sterility, 101, 78–86.
Bartel D P. 2009. MicroRNAs: Target recognition and regulatory functions. Cell, 136, 215–233.
Chen H, Fan Y, Xu W, Chen J, Xu C, Wei X, Fang D, Feng Y. 2016. miR-10b inhibits apoptosis and promotes proliferation and invasion of endometrial cancer cells via targeting HOXB3. Cancer Biotherapy and Radiopharmaceuticals, 31, 225–231.
Dai T, Vera Y, Salido E C, Yen P H. 2001. Characterization of the mouse Dazap1 gene encoding an RNA-binding protein that interacts with infertility factors DAZ and DAZL. BMC Genomics, 2, 6.
Eisenberg I, Kotaja N, Goldman-Wohl D, Imbar T. 2015. microRNA in human reproduction. Advances in Experimental Medicine and Biology, 888, 353–387.
Gerber J, Heinrich J, Brehm R. 2016. Blood-testis barrier and Sertoli cell function: Lessons from SCCx43KO mice. Reproduction, 151, R15–R27.
Hansen T B, Jensen T I, Clausen B H, Bramsen J B, Finsen B, Damgaard C K, Kjems J. 2013. Natural RNA circles function as efficient microRNA sponges. Nature, 495, 384–388.
Holt J E, Stanger S J, Nixon B, McLaughlin E A. 2016. Non-coding RNA in spermatogenesis and epididymal maturation. Advances in Experimental Medicine and Biology, 886, 95–120.
Hori T, Taguchi Y, Uesugi S, Kurihara Y. 2005. The RNA ligands for mouse proline-rich RNA-binding protein (mouse Prrp) contain two consensus sequences in separate loop structure. Nucleic Acids Research, 33, 190–200.
Hsu L C, Chen H Y, Lin Y W, Chu W C, Lin M J, Yan Y T, Yen P H. 2008. DAZAP1, an hnRNP protein, is required for normal growth and spermatogenesis in mice. RNA, 14, 1814–1822.
Hu P, Guan K, Feng Y, Ma C, Song H, Li Y, Xia X, Li J, Li F. 2017. miR-638 Inhibits immature Sertoli cell growth by indirectly inactivating PI3K/AKT pathway via SPAG1 gene. Cell Cycle, 16, 2290–2300.
Huang J, Sun C, Wang S, He Q, Li D. 2015. microRNA miR-10b inhibition reduces cell proliferation and promotes apoptosis in non-small cell lung cancer (NSCLC) cells. Molecular BioSystems, 11, 2051–2059.
Johnson L, Thompson Jr D L,Varner D D. 2008. Role of Sertoli cell number and function on regulation of spermatogenesis. Animal Reproduction Science, 105, 23–51.
Kurihara Y, Watanabe H, Kawaguchi A, Hori T, Mishiro K, Ono M, Sawada H, Uesugi S. 2004. Dynamic changes in intranuclear and subcellular localizations of mouse Prrp/DAZAP1 during spermatogenesis: The necessity of the C-terminal proline-rich region for nuclear import and localization. Archives of Histology and Cytology, 67, 325–333.
Li J, Liu X, Hu X, Tian G G, Ma W, Pei X, Wang Y, Wu J. 2017. MicroRNA-10b regulates the renewal of spermatogonial stem cells through Kruppel-like factor 4. Cell Biochemistry and Function, 35, 184–191.
Li Y, Li J, Fang C, Shi L, Tan J, Xiong Y, Bin F, Li C. 2016. Genome-wide differential expression of genes and small RNAs in testis of two different porcine breeds and at two different ages. Scientific Reports, 6, 26852.
Lian C, Sun B, Niu S, Yang R, Liu B, Lu C, Meng J, Qiu Z, Zhang L, Zhao Z. 2012. A comparative profile of the microRNA transcriptome in immature and mature porcine testes using Solexa deep sequencing. The FEBS Journal, 279, 964–975.
Liao C G, Kong L M, Zhou P, Yang X L, Huang J G, Zhang H L, Lu N. 2014. miR-10b is overexpressed in hepatocellular carcinoma and promotes cell proliferation, migration and invasion through RhoC, uPAR and MMPs. Journal of Translational Medicine, 12, 234.
Liao X, Ma C, Lu L, Zhang L, Luo X. 2017. Determination of dietary iron requirements by full expression of iron-containing cytochrome c oxidase in the heart of broilers from 22 to 42 d of age. The British Journal of Nutrition, 118, 493–499.
Liao X D, Wen Q, Zhang L Y, Lu L, Zhang L Y, Luo X G. 2018. Effect of dietary supplementation with flavonoid from Scutellaria baicalensis Georgi on growth performance, meat quality and antioxidative ability of broilers. Journal of Integrative Agriculture, 17, 1165–1170.
Lin Y T,Yen P H. 2006. A novel nucleocytoplasmic shuttling sequence of DAZAP1, a testis-abundant RNA-binding protein. RNA, 12, 1486–1493.
Lucas T F, Nascimento A R, Pisolato R, Pimenta M T, Lazari M F, Porto C S. 2014. Receptors and signaling pathways involved in proliferation and differentiation of Sertoli cells. Spermatogenesis, 4, e28138.
Luo L F, Hou C C, Yang W X. 2016. Small non-coding RNAs and their associated proteins in spermatogenesis. Gene, 578, 141–157.
Ma C, Song H, Guan K, Zhou J, Xia X, Li F. 2016a. Characterization of swine testicular cell line as immature porcine Sertoli cell line. In Vitro Cellular and Developmental Biology (Animal), 52, 427–433.
Ma C, Song H, Yu L, Guan K, Hu P, Li Y, Xia X, Li J, Jiang S, Li F. 2016b. miR-762 promotes porcine immature Sertoli cell growth via the ring finger protein 4 (RNF4) gene. Scientific Reports, 6, 32783.
Ma X Y, Liu S B, Lu L, Li S F, Xie J J, Zhang L Y, Zhang J H, Luo X G. 2014. Relative bioavailability of iron proteinate for broilers fed a casein-dextrose diet. Poultry Science, 93, 556–563.
Ma Y, Yang H Z, Xu L M, Huang Y R, Dai H L, Kang X N. 2015. Testosterone regulates the autophagic clearance of androgen binding protein in rat Sertoli cells. Scientific Reports, 5, 8894.
Mruk D D, Cheng C Y. 2015. The mammalian blood-testis barrier: Its biology and regulation. Endocrine Reviews, 36, 564–591.
Nixon B, Stanger S J, Mihalas B P, Reilly J N, Anderson A L, Tyagi S, Holt J E, McLaughlin E A. 2015. The microRNA signature of mouse spermatozoa is substantially modified during epididymal maturation. Biology of Reproduction, 93, 91.
Pan H A, Lin Y S, Lee K H, Huang J R, Lin Y H, Kuo P L. 2005. Expression patterns of the DAZ-associated protein DAZAP1 in rat and human ovaries. Fertility and Sterility, 84(Suppl. 2), 1089–1094.
Peng J Y, An X P, Fang F, Gao K X, Xin H Y, Han P, Bao L J, Ma H D, Cao B Y. 2016. MicroRNA-10b suppresses goat granulosa cell proliferation by targeting brain-derived neurotropic factor. Domestic Animal Endocrinology, 54, 60–67.
Ran M L, Chen B, Wu M S, Liu X C, He C Q, Yang A Q, Li Z, Xiang Y J, Li Z H, Zhang S W. 2015. Integrated analysis of miRNA and mRNA expression profiles in development of porcine testes. Rsc Advances, 5, 63439–63449.
Sharpe R M, McKinnell C, Kivlin C, Fisher J S. 2003. Proliferation and functional maturation of Sertoli cells, and their relevance to disorders of testis function in adulthood. Reproduction, 125, 769–784.
Smith R W, Anderson R C, Smith J W, Brook M, Richardson W A, Gray N K. 2011. DAZAP1, an RNA-binding protein required for development and spermatogenesis, can regulate mRNA translation. RNA, 17, 1282–1295.
Tremblay J J. 2015. Molecular regulation of steroidogenesis in endocrine Leydig cells. Steroids, 103, 3–10.
Vera Y, Dai T, Hikim A P, Lue Y, Salido E C, Swerdloff R S, Yen P H. 2002. Deleted in azoospermia associated protein 1 shuttles between nucleus and cytoplasm during normal germ cell maturation. Journal of Andrology, 23, 622–628.
Wu J, Zhu H, Song W, Li M, Liu C, Li N, Tang F, Mu H, Liao M, Li X, Guan W, Li X, Hua J. 2014. Identification of conservative microRNAs in Saanen dairy goat testis through deep sequencing. Reproduction in Domestic Animals, 49, 32–40.
Yao C, Sun M, Yuan Q, Niu M, Chen Z, Hou J, Wang H, Wen L, Liu Y, Li Z, He Z. 2016. MiRNA-133b promotes the proliferation of human Sertoli cells through targeting GLI3. Oncotarget, 7, 2201–2219.
Yin X, Ma T, Han R, Ding J, Zhang H, Han X, Li D. 2017. MiR-301b-3p/3584-5p enhances low-dose mono-n-butyl phthalate (MBP)-induced proliferation by targeting Rasd1 in Sertoli cells. Toxicology in Vitro (An International Journal Published in Association with BIBRA), 47, 79–88.
Yu X, Li Z, Chen G, Wu W K. 2015. MicroRNA-10b induces vascular muscle cell proliferation through Akt pathway by targeting TIP30. Current Vascular Pharmacology, 13, 679–686.
Yu X, Li Z, Shen J, Wu W K, Liang J, Weng X, Qiu G. 2013. MicroRNA-10b promotes nucleus pulposus cell proliferation through RhoC-Akt pathway by targeting HOXD10 in intervetebral disc degeneration. PLoS ONE, 8, e83080.
No related articles found!
No Suggested Reading articles found!