title: miR-34c works downstream of p53 leading to dairy goat male germline stem-cell (mGSCs) apoptosis.
abstract: OBJECTIVE
  Recent lines of evidence have indicated that miR-34c can play important roles in regulation of the cell cycle, cell senescence and apoptosis of mouse and human tumour cells, spermatogenesis, and male germ-cell apoptosis. However, there is little information on the effects of miR-34c on proliferation and apoptosis of livestock male germ cells. The dairy goat is a convenient domestic species for biological investigation and application. The purpose of this study was to investigate the effects of miR-34c on apoptosis and proliferation of dairy goat male germline stem cells (mGSCs), as well as to determine the relationship between p53 and miR-34c in this species.
  
  
  METHODS
  Morphological observation, miRNA in situ hybridisation (ISH), bromodeoxyuridine staining, flow cytometry, quantitative-RT-PCR (Q-RT-PCR) and western blotting were utilized to ascertain apoptosis and proliferation of mGSCs, through transfection of miR-34c mimics (miR-34c), miR-34c inhibitor (anti-miR-34c), miR-34c mimics and inhibitors co-transfected (mixture) compared to control groups.
  
  
  RESULTS
  Results manifested that miR-34c over-expression promoted mGSCs apoptosis and suppressed their proliferation. Simultaneously, a variety of apoptosis-related gene expression was increased while some proliferation-related genes were downregulated. Accordingly, miR-34c promoted apoptosis in mGSCs and reduced their proliferation; moreover, expression of miR-34c was p53-dependent.
  
  
  CONCLUSIONS
  This study is the first to provide a model for study of miRNAs and mechanisms of proliferation and apoptosis in male dairy goat germ cells.
authors.affiliation: College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry, Northwest A&F University, Yangling, Shaanxi, 712100, China.
authors.forename: M
authors.initials: M
authors.lastname: Li
authors.forename: M
authors.initials: M
authors.lastname: Yu
authors.forename: C
authors.initials: C
authors.lastname: Liu
authors.forename: H
authors.initials: H
authors.lastname: Zhu
authors.forename: X
authors.initials: X
authors.lastname: He
authors.forename: S
authors.initials: S
authors.lastname: Peng
authors.forename: J
authors.initials: J
authors.lastname: Hua
mesh_terms: D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D001973:Bromodeoxyuridine / Q000378:metabolism; D049109:Cell Proliferation; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000494:pharmacology; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D006041:Goats / Q000254:growth & development / Q000378:metabolism*; D017403:In Situ Hybridization; D008297:Male; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D013094:Spermatozoa / Q000378:metabolism / Q000473:pathology; D013234:Stem Cells / Q000378:metabolism / Q000473:pathology*; D013737:Testis / Q000378:metabolism / Q000473:pathology; D013997:Time Factors; D015533:Transcriptional Activation; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
chemical_list: D035683:MicroRNAs; D016159:Tumor Suppressor Protein p53; D004317:Doxorubicin; D001973:Bromodeoxyuridine